This invention relates to substituted aromatic tricyclic compounds containing nicotinonitrile rings as well as the pharmaceutically acceptable salts thereof The compounds of the present invention inhibit the action of certain protein kinases, thereby inhibiting the abnormal growth of particular cell types. The compounds of this invention are therefore useful for the treatment or inhibition of certain diseases that are the result of deregulation of these protein kinases. The compounds of this invention are anti-cancer agents and are useful for the treatment or inhibition of cancer in mammals. In addition, the compounds of this invention are useful for the treatment and inhibition of polycystic kidney disease and colonic polyps.
Protein kinases are a class of enzymes that catalyze the transfer of a phosphate group from ATP to a tyrosine, serine, threonine, or histidine residue located on a protein substrate. Protein kinases clearly play a role in normal cell growth. Many of the growth factor receptor proteins function as kinases and it is by this process that they effect signaling. The interaction of growth factors with these receptors is a necessary event in normal regulation of cell growth. However, under certain conditions, as a result of either mutation or over expression, these receptors can become deregulated; the result of which is uncontrolled cell proliferation which can lead to tumor growth and ultimately to the disease known as cancer [Wilks, A. F., Adv. Cancer Res., 60, 43 (1993) and Parsons, J. T.; Parsons, S. J., Important Advances in Oncology, DeVita, V. T. Ed., J. B. Lippincott Co., Phila., 3 (1993)]. Among the growth factor receptor kinases and their proto-oncogenes that have been identified and which are targets of the compounds of this invention are the epidermal growth factor receptor kinase (EGF-R kinase, the protein product of the erbB oncogene), and the product produced by the erbB-2 (also referred to as the neu or HER2) oncogene. Since the phosphorylation event is a necessary signal for cell division to occur and since overexpressed or mutated kinases have been associated with cancer, an inhibitor of this event, a protein tyrosine kinase inhibitor, will have therapeutic value for the treatment of cancer and other diseases characterized by uncontrolled or abnormal cell growth. For example, over expression of the receptor kinase product of the erbB-2 oncogene has been associated with human breast and ovarian cancers [Slamon, D. J. et al., Science, 244, 707 (1989) and Science, 235 , 177 (1987)]. Deregulation of EGF-R kinase has been associated with epidermoid tumors [Reiss, M., et al., Cancer Res., 51, 6254 (1991)], breast tumors [Macias, A. et al., Anticancer Res., 7, 459 (1987)], and tumors involving other major organs [Gullick, W. J., Brit. Med. Bull., 47, 87 (1991)]. Because of the importance of the role played by deregulated receptor kinases in the pathogenesis of cancer, many recent studies have dealt with the development of specific PTK inhibitors as potential anti-cancer therapeutic agents [some recent reviews: Traxler, P., Exp. Opin. Ther. Patents, 8, 1599 (1998) and Bridges, A. J., Emerging Drugs, 3, 279 (1998)].
It is also known that deregulation of EGF receptors is a factor in the growth of epithelial cysts in the disease described as polycystic kidney disease [Du, J., Wilson, P. D., Amer. J. Physiol., 269 (2 Pt 1), 487 (1995); Nauta, J., et al., Pediatric Research, 37(6), 755 (1995); Gattone, V. H. et al., Developmental. Biology, 169(2), 504 (1995); Wilson, P. D. et al., Eur. J. Cell Biol., 61(1), 131, (1993)]. The compounds of this invention, which inhibit the catalytic function of the EGF receptors, are consequently useful for the treatment of this disease.
The mitogen-activated protein kinase (MAPK) pathway is a major pathway in the cellular signal transduction cascade from growth factors to the cell nucleus. The pathway involves kinases at two levels: MAP kinase kinases (MAPKK), and their substrates MAP kinases (MAPK). There are different isoforms in the MAP kinase family. (For review, see Seger, R.; Krebs, E. G., FASEB, 9, 726, (1995).) The compounds of this invention can inhibit the action of two of these kinases: MEK, a MAP kinase kinase, and its substrate ERK, a MAP kinase. MEK is activated by phosphorylation on two serine residues by upstream kinases such as members of the raf family. When activated, MEK catalyzes phosphorylation on a threonine and a tyrosine residue of ERK. The activated ERK then phosphorylates and activates transcription factors in the nucleus, such as fos and jun, or other cellular targets with PXT/SP sequences. ERK, a p42 MAPK, is found to be essential for cell proliferation and differentiation. Over-expression and/or over-activation of MEK or ERK has been found to be associated with various human cancers [For example, Sivaraman, V. S.; Wang, H-Y.; Nuovo, G. J. Malbon, C. C. J. Clin. Invest., 99, 1478 (1997)]. It has been demonstrated that inhibition of MEK prevents activation of ERK and subsequent activation of ERK substrates in cells, resulting in inhibition of cell growth stimulation and reversal of the phenotype of ras-transformed cells [Dudley, D. T.; Pang, L.; Decker, S. J.; Bridges, A. J.; Saltiel, A. R., Proc. Nat. Acad. Sci., 92, 7686, (1995)]. Since, as demonstrated below, the compounds of this invention can inhibit the coupled action of MEK and ERK, they are useful for the treatment of diseases such as cancer which are characterized by uncontrolled cell proliferation and which, at least in part, depend on the MAPK pathway.
As mentioned above, members of the raf family of kinases phosphorylate serine residues on MEK. There are three serine/threonine kinase members of the raf family known as a-raf, b-raf and c-raf. While mutations in the raf genes are rare in human cancers, c-raf is activated by the ras oncogene which is mutated in a wide number of human tumors. Therefore inhibition of the kinase activity of c-raf may provide a way to prevent ras mediated tumor growth [Campbell, S. L., Oncogene, 17, 1395 (1998)].
The Src family of cytoplasmic protein tyrosine kinases consists of at least eight members (Src, Fyn, Lyn, Yes, Lck, Fgr, Hck and Blk) that participate in a variety of signaling pathways [Schwartzberg, P. L., Oncogene, 17, 1463-1468, (1998)]. The prototypical member of this tyrosine kinase family is p60src (Src). Src is involved in proliferation and migration responses in many cell types. In limited studies, Src activity has been shown to be elevated in breast, colon (xcx9c90%), pancreatic ( greater than 90%) and liver ( greater than 90%) tumors. Greatly increased Src activity is also associated with metastasis ( greater than 90%) and poor prognosis. Antisense Src message impedes growth of colon tumor cells in nude mice [Staley et al., Cell Growth and Differentiation., 8, 269-74, (1997)], suggesting that Src inhibitors should slow tumor growth. In addition to its role in cell proliferation, Src also acts in stress response pathways, including the hypoxia response, and nude mice studies with colon tumor cells expressing antisense Src message have reduced vascularization [Ellis, et al., J. Biol. Chem., 273, 1052-7 (1998)], which suggests that Src inhibitors would be anti-angiogenic as well as anti-proliferative.
In addition to its role in cancer, Src also appears to play a role in osteoporosis. Mice genetically engineered to be deficient in src production were found to exhibit osteopetrosis, the failure to resorb bone [Soriano, P., Cell, 64, 693 (1991); Boyce, B. F., J. Clin., Invest., 90, 1622 (1992)]. This defect was characterized by a lack of osteoclast activity. Since osteoclasts normally express high levels of Src, inhibition of Src kinase activity may be useful in the treatment of osteoporosis [Missbach, M., Bone, 24, 437 (1999)].
In addition to EGFr, there are several other RTKs including FGFr, the receptor for fibroblast growth factor (FGF); flk-1, also known as KDR, and flt-1, the receptors for vascular endothelial growth factor (VEGF); and PDGFr, the receptor for platelet derived growth factor (PDGF). The formation of new blood vessels, a process known as angiogenesis, is essential for tumor growth. Two natural angiogenesis inhibitors, angiostatin and endostatin, dramatically inhibited the growth of a variety of solid tumors. [O""Reilly, M. S., Cell, 79, 315 (1994); O""Reilly, M. S., Nature Medicine, 2, 689 (1996); O""Reilly, M. S., Cell, 88, 277 (1997)]. Since FGF and VEGF are known to stimulate angiogenesis, inhibition of the kinase activity of their receptors should block the angiogenic effects of these growth factors. In addition, the receptor tyrosine kinases tie-1 and tie-2 also play a key role in angiogenesis [Sato, T. N., Nature, 376, 70 (1995)]. Compounds of the invention that inhibit the kinase activity of FGFr, flk-1, flt-1, tie-1 or tie-2 may inhibit tumor growth by their effect on angiogenesis. Normal angiogenesis is required in many physiological conditions such as wound healing, female reproduction and fetal development. Abnormal or pathological angiogenesis has been implicated in neoplastic diseases including solid tumor growth, metastasis, and Karposi""s sarcoma; various eye diseases including diabetic retinopathy, and macular degeneration; inflammatory conditions including rheumatoid arthritis, and osteoarthritis; skin diseases including psoriasis, eczema and scleroderma; as well as ulcerative colitis and childhood haemangiomas [Toi, M. et al., Breast Cancer Res. And Treat., 36, 192-204 (1995); Folkman, J., Nature Medicine, 1, 27-3 1 (1995); Jackson, J. R. et al., FASEB J., 11, 457-465 (1997)]. Inhibition of VEGF function has been shown to inhibit disease progression in tumors [Borgstrom, P. et al., Cancer Res., 56, 4032-4039 (1996); Kim, J. K. et al., Nature, 362, 841-844 (1993)] and retinal neovascularization [Aiello, L. P. et al., Proc. Nat. Acad. Sci., 92, 10457-10461 (1995)] as well as vascular dysfunction mediated by glucose in models of diabetes [Tilton, R. G. et al., J. Clin. Invest., 99, 2192-2202 (1997)].
PDGF is a potent growth factor and chemoattractant for smooth muscle cells (SMCs) and the renarrowing of coronary arteries following angioplasty is due in part to the enhanced proliferation of SMCs in response to increased levels of PDGF. Therefore, compounds that inhibit the kinase activity of PDGFr may be useful in the treatment of restenosis. In addition, since PDGF and PDGFr are overexpressed in several types of human gliomas, small molecules capable of suppressing PDGFr activity, have potential utility as anticancer therapeutics [Nister, M., J. Biol. Chem. 266, 16755 (1991); Strawn, L. M., J. Biol. Chem. 269, 21215 (1994)].
In accordance with the present invention, the tricyclic ring systems described herein will be numbered as indicated in the representative formulas below (where U=N or O or S): 
No fully aromatic fused tricyclic compounds containing nicotinonitrile rings have been reported that have biological activity as inhibitors of protein tyrosine kinases. 3-Cyanoquinoline derivatives described in WO-9843960 have been disclosed as inhibitors of tyrosine kinase. A 3-cyanoquinoline with a 4-(2-methyl anilino) substituent having gastric (H+/K+)-ATPase inhibitory activity at high concentrations has been described [Ife R. J., et al., J. Med. Chem., 35(18), 3413 (1992)]. However, there are no references to any fully aromatic tricyclic compounds containing nicotinonitrile rings in the above publications.
In WO-9713760, a series of fused tricyclic compounds containing pyridine rings (and pyrimidines) that are reported to be inhibitors of protein tyrosine kinases is disclosed. However, it is specified that the position meta to the pyridine nitrogen bears a hydrogen atom only. No compounds possessing cyano substituents at this position are claimed. In two patents: AU 8767450 and U.S. Pat. No. 4,952,584, 4-amino-9H-pyrido-(2,3-b)-indole-3-carboxylic acid derivatives are disclosed as anxiolytic and antidepressant agents. No corresponding 3-cyano substituents are claimed. In EP 755934, fused tricyclic compounds containing nicotinonitrile rings are disclosed as endothelin receptor antagonists. However, these derivatives do not have the unique combination of substituents contained in the compounds of the present invention. In particular, it is specified that these compounds possess aromatic substituents directly attached to the position para to the pyridine nitrogen. Such substituents are not claimed in the present invention. Similarly, a series of compounds claimed in WO 9705137 do include tricyclics containing nicotinonitrile rings, but with hydrogen or simple alkyl chains attached to the position para to the pyridine nitrogen. Such substituents are not claimed in the present invention. Several patents exist which disclose substituted quinoline compounds as tyrosine kinase inhibitors, none of which possess the 3-cyano substituent: 1. An international patent WO-9813350 describing 3-fluoroquinoline and quinoline tyrosine kinase inhibitors. 2. WO-9609294 discloses inhibitors of protein tyrosine kinases that include 4-anilino quinolines with a large variety of substituents on positions 5-8 but which must also have a hydrogen atom at position 3. 3. U.S. Pat. No. 5,480,883 discloses quinoline derivatives that are inhibitors of protein tyrosine kinases but these derivatives do not have the unique combination of substituents, including the 3-cyano group, contained in the compounds of the present invention.
In addition to the above-mentioned compounds, certain tricyclics containing pyrimidine rings are known to be inhibitors of protein tyrosine kinases. WO-9749688, WO-9519970, U.S. Pat. No. 5,679,683 and the previously-mentioned WO-9713760 disclose a variety of tricyclic heterocycles which are tyrosine kinase inhibitors. Other patent applications WO-9802434, WO-9730044 and EP-837063 describe quinazolines substituted at positions 5 to 8 with one or more optionally substituted 5- or 6-membered heterocyclic rings.
In addition to the aforementioned patent applications, a number of publications describe fused tricyclics containing 4-anilinopyrimidine rings: Rewcastle G. W., et. al., J. Med. Chem., 39, 918 (1996); Bencteux, E., et. al., J. Heterocycl. Chem., 34, 1375, (1997); Palmer B. D., et. al. J. Med. Chem., 40, 1519 (1997); and Zhou, H., et. al., Book of Abstracts, 210th ACS National Meeting, Chicago, Ill., Aug. 20-24 (1995), Issue Pt. 2, MEDI-017. There are no publications that describe fused tricyclic tricyclic compounds containing nicotinonitrile rings as PTK inhibitors.
The present invention relates to certain protein kinase inhibitors of formula 1 having the structure: 
wherein:
Ar is cycloalkyl of 3 to 7 carbon atoms, which may be optionally substituted with one or more alkyl of 1 to 6 carbon atoms; or
Ar is a pyridinyl, pyrimidinyl, or phenyl ring; wherein the pyridinyl, pyrimidinyl, or phenyl ring may be optionally mono-, di-, or tri-substituted with substituents selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, carboxyalkyl of 2-7 carbon atoms, carboalkoxyalkyl of 3-8 carbon atoms, aminoalkyl of 1-5 carbon atoms, N-alkylaminoalkyl of 2-9 carbon atoms, N,N-dialkylaminoalkyl of 3-10 carbon atoms, N-alkylaminoalkoxy of 3-9 carbon atoms, N,N-dialkylaminoalkoxy of 4-10 carbon atoms, mercapto, methylmercapto and benzoylamino; or
Ar is a bicyclic aryl or bicyclic heteroaryl ring system of 8 to 12 atoms where the bicyclic heteroaryl ring may contain 1 to 4 heteroatoms selected from N, O, and S wherein the bicyclic aryl or bicyclic heteroaryl ring may be optionally mono- di-, tri, or tetra-substituted with substituent(s) independently selected from the group consisting of halogen, oxo, thiocarbonyl, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, phenoxy, phenyl, thiophenoxy, benzoyl, benzyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms, phenylamino, benzylamino, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, carboxyalkyl of 2-7 carbon atoms, carboalkoxyalkyl of 3-8 carbon atoms, aminoalkyl of 1-5 carbon atoms, N-alkylaminoalkyl of 2-9 carbon atoms, N,N-dialkylaminoalkyl of 3-10 carbon atoms, N-alkylaminoalkoxy of 3-9 carbon atoms, N,N-dialkylaminoalkoxy of 4-10 carbon atoms, mercapto, methylmercapto, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
Axe2x80x2 is a pyridinyl, pyrimidinyl, or phenyl ring; wherein the pyridinyl, pyrimidinyl, or phenyl ring may be optionally mono- or di-substituted with a substituent(s) independently selected from the group consisting of alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halogen, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, phenoxy, phenyl, thiophenoxy, benzoyl, benzyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms, phenylamino, benzylamino, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, carboxyalkyl of 2-7 carbon atoms, carboalkoxyalkyl of 3-8 carbon atoms, aminoalkyl of 1-5 carbon atoms, N-alkylaminoalkyl of 2-9 carbon atoms, N,N-dialkylaminoalkyl of 3-10 carbon atoms, N-alkylaminoalkoxy of 3-9 carbon atoms, N,N-dialkylaminoalkoxy of 4-10 carbon atoms, mercapto, methylmercapto, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino;
T is substituted on Axe2x80x2 at carbon and is xe2x80x94NH(CH2)mxe2x80x94, xe2x80x94O(CH2)mxe2x80x94, xe2x80x94S(CH2)mxe2x80x94, xe2x80x94NR(CH2)mxe2x80x94, xe2x80x94(CH2)mxe2x80x94, xe2x80x94(CH2)mNHxe2x80x94, xe2x80x94(CH2)mOxe2x80x94, xe2x80x94(CH2)mSxe2x80x94, xe2x80x94SO(CH2)mxe2x80x94, xe2x80x94SO2(CH2)mxe2x80x94, xe2x80x94CO(CH2)mxe2x80x94, xe2x80x94(CH2)mCOxe2x80x94, xe2x80x94(CH2)mSOxe2x80x94, xe2x80x94(CH2)mSO2xe2x80x94 or xe2x80x94(CH2)mNRxe2x80x94;
L is a phenyl ring that is optionally substituted with one, two, or three substituent(s) independently selected from the group consisting of alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halogen, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, phenoxy, phenyl, thiophenoxy, benzoyl, benzyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms, phenylamino, benzylamino, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, carboxyalkyl of 2-7 carbon atoms, carboalkoxyalkyl of 3-8 carbon atoms, aminoalkyl of 1-5 carbon atoms, N-alkylaminoalkyl of 2-9 carbon atoms, N,N-dialkylaminoalkyl of 3-10 carbon atoms, N-alkylaminoalkoxy of 3-9 carbon atoms, N,N-dialkylaminoalkoxy of 4-10 carbon atoms, mercapto, methylmercapto, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
L is a 5- or 6-membered heteroaryl ring where the heteroaryl ring contains 1 to 3 heteroatoms selected from N, O, and S and where the heteroaryl ring may be optionally mono- or di-substituted with substituent(s) selected from the group consisting of halogen, oxo, thiocarbonyl, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, phenoxy, phenyl, thiophenoxy, benzoyl, benzyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms, phenylamino, benzylamino, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, carboxyalkyl of 2-7 carbon atoms, carboalkoxyalkyl of 3-8 carbon atoms, aminoalkyl of 1-5 carbon atoms, N-alkylaminoalkyl of 2-9 carbon atoms, N,N-dialkylaminoalkyl of 3-10 carbon atoms, N-alkylaminoalkoxy of 3-9 carbon atoms, N,N-dialkylaminoalkoxy of 4-10 carbon atoms, mercapto, methylmercapto, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino;
m is 0-3;
n is 0-1;
X is NH, O, S, or NR;
R is alkyl of 1-6 carbon atoms;
Y and Z are both carbon or N; the ring structure of formula 1 then being a fused 5,6,6 or 6,6,6 tricycle; or one of Y and Z is N, O or S, and the other is a bond between the two end rings; the ring structure of formula 1 then being a fused 5,5,6 or 6,5,6 tricycle; or one of Y or Z is N with the other being carbon; the ring structure of formula 1 then being a fused 5,6,6 or 6,6,6 tricycle; 
A and D are each, independently, carbon, N, O, or S;
B is carbon or N;
the dashed line indicates an optional double bond;
R1, R2, R3, and R4 are each, independently, not present, hydrogen, halogen, hydroxy, amino, hydroxyamino, trifluoromethyl, trifluoromethoxy, mercapto, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, alkynyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, alkenylsulfonamido of 2-6 carbon atoms, alkynylsulfonamido of 2-6 carbon atoms, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, alkenoyl of 3-7 carbon atoms, N-alkyl-N-alkenylamino of 4 to 12 carbon atoms, N,N-dialkenylamino of 6-12 carbon atoms, phenylamino, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, alkanoyloxy of 2-7 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, dialkylamino of 2 to 12 carbon atoms, alkanoyloxymethyl group of 2-7 carbon atoms, alkenoyloxymethyl group of 2-7 carbon atoms, alkynoyloxymethyl group of 2-7 carbon atoms, azido, benzoyl, carboxyalkyl of 2-7 carbons, carboalkoxyalkyl of 3-8 carbon atoms, 
R5 is independently hydrogen, alkyl of 1-6 carbon atoms, aminoalkyl of 1-6 carbon atoms, N-alkylaminoalkyl of 2-9 carbon atoms, N,N-dialkylaminoalkyl of 3-12 carbon atoms, N-cycloalkylaminoalkyl of 4-12 carbon atoms, N-cycloalkyl-N-alkylaminoalkyl of 5-18 carbon atoms, N,N-dicycloalkylaminoalkyl of 7-18 carbon atoms, morpholino-N-alkyl wherein the alkyl group is 1-6 carbon atoms, piperidino-N-alkyl wherein the alkyl group is 1-6 carbon atoms, N-alkyl-piperazino-N-alkyl wherein either alkyl group is 1-6 carbon atoms, azacycloalkyl-N-alkyl of 3-11 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, alkoxyalkyl of 2-8 carbon atoms, or phenyl;
V is (CH2)m, O, S, or NR6;
R7 is NR6R6, OR6, J, N(R6)3+, or NR6(OR6);
M is NR6, O, S, Nxe2x80x94[(C(R6)2)pNR6R6], or Nxe2x80x94[(C(R6)2)pxe2x80x94OR6];
W is NR6, O, S, or is a bond;
Het is a heterocycle selected from the group consisting of morpholine, thiomorpholine, thiomorpholine S-oxide, thiomorpholine S,S-dioxide, piperidine, pyrrolidine, aziridine, pyridine, imidazole, 1,2,3-triazole, 1,2,4-triazole, thiazole, thiazolidine, tetrazole, piperazine, furan, thiophene, tetrahydrothiophene, tetrahydrofuran, dioxane, 1,3-dioxolane pyrrole, and tetrahydropyran; wherein the heterocycle is optionally mono- or di-substituted on carbon or nitrogen with R6; optionally mono- or di-substituted on carbon with hydroxy, xe2x80x94N(R6)2, or xe2x80x94OR6; optionally mono or di-substituted on carbon with the mono-valent radicals xe2x80x94(C(R6)2)sOR6 or xe2x80x94[(C(R6)2)sN(R6)2]; or optionally mono or di-substituted on a saturated carbon with divalent radicals xe2x95x90O or xe2x80x94O(C(R6)2)sOxe2x80x94;
Ph is a phenyl ring optionally mono-, di- or tri-substituted with halogen, alkyl of 1-6 carbon atoms, trifluoromethyl, nitro, cyano, azido, halomethyl, carboxyl, alkoxycarbonyl, alkylthio, mercapto, mercaptomethyl, xe2x80x94N(R6)2, xe2x80x94OR6, xe2x80x94(C(R6)2)sOR6, xe2x80x94[(C(R6)2)sN(R6)2], or xe2x80x94(C(R6)2)kHet;
R6 is hydrogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, cycloalkyl of 1-6 carbon atoms, alkanoyl of 2-7 carbon atoms, carbamoylalkyl of 2-7 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, hydroxycycloalkyl of 3-6 carbon atoms, or carboxyalkyl of 2-7 carbon atoms; or
R6 is phenyl optionally mono-, di-, or tri-substituted with substituent(s) independently selected from halogen, alkoxy of 1-6 carbon atoms, trifluoromethyl, amino, alkylamino of 1-3 carbon atoms, dialkylamino of 2-6 carbon atoms, nitro, cyano, azido, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, carboxyl, alkoxycarbonyl of 2-7 carbon atoms, phenoxy, phenyl, thiophenoxy, benzoyl, benzyl, phenylamino, benzylamino; alkanoylamino of 1-6 carbon atoms or alkyl of 1-6 carbon atoms;
R8 and R9 are each, independently, xe2x80x94[(C(R6)2)rNR6R6], and xe2x80x94[(C(R6)2)rOR6];
J is independently hydrogen, chlorine, fluorine, or bromine;                               g          =                      1            -            6                          ;                                          k          =                      0            -            4                          ;                                          p          =                      2            -            4                          ;                                          q          =                      0            -            4                          ;                                          r          =                      1            -            4                          ;                                          s          =                      1            -            6                          ;            
or a pharmaceutically acceptable salt thereof;
provided that when 
at least one of the bonds between A and B or B and D must be a double bond, with the other being a single bond;
at least one of A, B, and D are not carbon;
only one of A, B, or D can be O or S;
when A, B, or D is O or S, the adjacent atoms must be carbon;
provided that when R5 is bound to a nitrogen atom, the resulting structures do not include xe2x80x94Nxe2x80x94Cxe2x80x94Nxe2x80x94 or xe2x80x94Oxe2x80x94Cxe2x80x94Nxe2x80x94 radicals; and when R5 is bound to an oxygen atom, the resulting structures do not include an xe2x80x94Nxe2x80x94Cxe2x80x94Oxe2x80x94 radical;
provided that when R6 is alkenyl of 2-7 carbon atoms or alkynyl of 2-7 carbon atoms, the alkenyl or alkynyl moieties are bound to a nitrogen or oxygen atom through a saturated carbon atom in the alkenyl or alkynyl chain;
provided that when V is NR6 and R7 is NR6R6, N(R6)3+, or NR6(OR6), then g=2-6;
provided that when M is O or S and R7 is OR6, then p=1-4;
provided that when V is NR6, O, S, then k=2-4;
provided that when V is O or S and M or W is O or S, then k=1-4
provided that when W is not a bond with Het bonded through a nitrogen atom then q=2-4; and
finally provided when W is a bond with Het bonded through a nitrogen atom and V is O or NR6 or S, then k=2-4.
The present invention also relates to a method for making compounds of formula 1 and methods of using the compounds of formula 1.
The present invention relates to substituted aromatic tricyclic compounds containing nicotinonitrile rings of formula 1 above as well as the pharmaceutically acceptable salts thereof. The compounds of the present invention inhibit the action of certain protein kinases, thereby inhibiting the abnormal growth of particular cell types. The compounds of this invention are therefore useful for the treatment or inhibition of certain diseases that are the result of deregulation of these protein kinases. The compounds of this invention are anti-cancer agents and are useful for the treatment or inhibition of cancer in mammals. In addition, the compounds of this invention are useful for the treatment and inhibition of polycystic kidney disease and colonic polyps.
The pharmaceutically acceptable salts are any conventionally known salts useful in the pharmaceutical industry including those derived from such organic and inorganic acids such as: acetic, lactic, citric, tartaric, succinic, maleic, malonic, gluconic, hydrochloric, hydrobromic, phosphoric, nitric, sulfuric, methanesulfonic, and similarly known acceptable acids.
In the present application in those cases in which a substituent, moiety, or group is di-, tri-, and/or tetra-substituted, it is understood that the 2, 3, and/or 4 substituents on the substituent, moiety, or group may be the same or different.
It is understood by one skilled in the art that the heteroaryl or bicyclic heteroaryl rings of the compounds of Formula I do not contain Oxe2x80x94O, Sxe2x80x94S, or Sxe2x80x94O bonds, as they would be unstable. Preferred bicyclic aryl or bicyclic heteroaryl ring systems include naphthalene, tetralin, indan, 1-indanone, 1,2,3,4-tetrahydroquinoline, naphthyridine, benzofuran, 3-oxo-1,3-dihydroisobenzofuran, benzothiophene, 1,1-dioxobenzothiophene, indole, indoline 1,3-dioxo-2,3-dihydro-1H-isoindole, benzotriazole, 1H-indazole, indoline, indazole, 1,3-benzodioxole, benzoxazole, purine, phthalimide, coumarin, chromone, quinoline, terahydroquinoline, isoquinoline, benzimidazole, quinazoline, pyrido[2,3-b]pyridine, pyrido[3,4-b]pyrazine, pyrido[3,2-c]pyridazine, pyrido[3,4-b]pyridine, 1H-pyrazole[3,4-d]pyrimidine, 1,4-benzodioxane, pteridine, 2(1H)-quinolone, 1(2H)-isoquinolone, 2-oxo-2,3-dihydrobenzthiazole, 1,2-methylenedioxybenzene, 2-oxindole, 1,4-benzisoxazine, benzothiazole, quinoxaline, quinoline-N-oxide, isoquinoline-N-oxide, quinoxaline-N-oxide, quinazoline-N-oxide, benzoazine, phthalazine, 1,4-dioxo-1,2,3,4-tetrahydrophthalazine, 2-oxo-1,2-dihydroquinoline, 2,4-dioxo-1,4-dihydro-2H-benzo[d][1,3]oxazine, 2,5-dioxo-2,3,4,5-tetrahydro-1H-benzo[e][1,4]diazepine, or cinnoline.
When L is a 5 or 6-membered heteroaryl ring, preferred heteroaryl rings are pyridine, pyrimidine, imidazole, thiazole, thiazolidine, pyrrole, furan, thiophene, oxazole, or 1,2,4-triazole.
Either or both rings of the bicyclic aryl or bicyclic heteroaryl group may be fully unsaturated, partially saturated, or fully saturated. An oxo substituent on the bicyclic aryl or bicyclic heteroaryl moiety means that one of the carbon atoms has a carbonyl group. A thiocarbonyl substituent on the bicyclic aryl or bicyclic heteroaryl moiety means that one of the carbon atoms has a thiocarbonyl group.
When L is a 5 or 6-membered heteroaryl ring, it may be fully unsaturated, partially saturated, or fully saturated. The heteroaryl ring can be bound to Axe2x80x2 via carbon or nitrogen. An oxo substituent on the heteroaryl ring means that one of the carbon atoms has a carbonyl group. A thio substituent on the heteroaryl ring means that one of the carbon atoms has a thiocarbonyl group.
The alkyl portion of the alkyl, alkoxy, alkanoyloxy, alkoxymethyl, alkanoyloxymethyl, alkylsulphinyl, alkylsulfonyl, alkylsulfonamido, alkoxycarbonyl, carboxyalkyl, carboalkoxyalkyl, alkanoylamino, N-alkylcarbamoyl, N,N-dialkylcarbamoyl, N-alkylaminoalkoxy and N,N-dialkylaminoalkoxy include both straight chain as well as branched carbon chains. The cycloalkyl portions of cycloalkyl, N-cycloalkylamino, N-cycloalkyl-N-alkylaminoalkyl, N,N-dicycloalkylaminoalkyl, cycloalkylthio and azacycloalkyl substituents include both simple carbocycles as well as carbocycles containing alkyl substituents. The alkenyl portion of the alkenyl, alkenyloxy, alkenylsulfonamido, substituents include both straight chain as well as branched carbon chains and one or more sites of unsaturation and all possible configurational isomers. The alkynyl portion of the alkynyl, alkynylsulfonamido, alkynyloxy, substituents include both straight chain as well as branched carbon chains and one or more sites of unsaturation. Carboxy is defined as a xe2x80x94CO2H radical. Alkoxycarbonyl of 2-7 carbon atoms is defined as a xe2x80x94CO2Rxe2x80x3 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms. Carboxyalkyl is defined as a HO2Cxe2x80x94Rxe2x80x2xe2x80x3xe2x80x94 radical where Rxe2x80x2xe2x80x3 is a divalent alkyl radical of 1-6 carbon atoms. Carboalkoxyalkyl is defined as a Rxe2x80x3O2Cxe2x80x94Rxe2x80x2xe2x80x3xe2x80x94 radical where Rxe2x80x2xe2x80x3 is a divalent alkyl radical and where Rxe2x80x3 and Rxe2x80x2xe2x80x3 may be the same or different, and together have 2-7 carbon atoms. Alkanoyl is defined as a xe2x80x94CORxe2x80x3 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms. Alkenoyl is defined as a xe2x80x94CORxe2x80x3 radical, where Rxe2x80x3 is an alkenyl radical of 2-6 carbon atoms. Alkanoyloxy is defined as a xe2x80x94OCORxe2x80x3 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms. Alkanoyloxymethyl is defined as Rxe2x80x3CO2CH2xe2x80x94 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms. Alkoxymethyl is defined as Rxe2x80x3OCH2xe2x80x94 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms. Alkylsulphinyl is defined as Rxe2x80x3SOxe2x80x94 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms. Alkylsulfonyl is defined as Rxe2x80x3 SO2xe2x80x94 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms. Alkylsulfonamido, alkenylsulfonamido, alkynylsulfonamido are defined as Rxe2x80x3SO2NHxe2x80x94 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms, an alkenyl radical of 2-6 carbon atoms, or an alkynyl radical of 2-6 carbon atoms, respectively. N-alkylcarbamoyl is defined as Rxe2x80x3NHCOxe2x80x94 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms. N,N-dialkylcarbamoyl is defined as Rxe2x80x3Rxe2x80x2NCOxe2x80x94 radical, where Rxe2x80x3 is an alkyl radical of 1-6 carbon atoms, Rxe2x80x2 is an alkyl radical of 1-6 carbon atoms and Rxe2x80x2 and Rxe2x80x3 may be the same or different.
Het is a heterocycle, as defined above which in some cases when Het is substituted with xe2x95x90O (carbonyl), the carbonyl group can be hydrated. Het may be bonded to W when q=0 via a carbon atom on the heterocyclic ring, or when Het is a nitrogen containing heterocycle which also contains a saturated carbon-nitrogen bond, such heterocycle may be bonded to carbon, via the nitrogen when W is a bond. When q=0 and Het is a nitrogen containing heterocycle which also contains an unsaturated carbon-nitrogen bond, that nitrogen atom of the heterocycle may be bonded to carbon when W is a bond and the resulting heterocycle will bear a positive charge. When Het is substituted with R6, such substitution may be on a ring carbon, or in the case of a nitrogen containing heterocycle, which also contains a saturated carbon-nitrogen, such nitrogen may be substituted with R6 or in the case of a nitrogen containing heterocycle, which also contains an unsaturated carbon-nitrogen, such nitrogen may be substituted with R6 in which case the heterocycle will bear a positive charge. Preferred heterocycles include pyridine, 2,6-disubstituted morpholine, 2,5-disubstituted thiomorpholine, 2-substituted imidazole, substituted thiazole, N-substituted imidazole, N-subsitituted 1,4-piperazine, N-subsitituted piperidine, and N-substituted pyrrolidine.
The compounds of this invention may contain one or more asymmetric carbons atoms; in such cases, the compounds of this invention include the individual diasteromers, the racemates, and the individual R and S enantiomers thereof. Some of the compounds of this invention may contain one or more double bonds; in such cases, the compounds of this invention include each of the possible configurational isomers as well as mixtures of these isomers.
Preferred compounds of the invention are selected from: 
wherein Ar, R1, R2, R3 and R4 are as defined above.
More preferred compounds of this invention are described below. Except as otherwise indicated below, the substituents are as defined above.
A. Compounds according to the formula 1, having the structure 
or a pharmaceutically acceptable salt thereof.
B. Compounds according to formula 1, having the structure 
X is selected from NH, sulfur or oxygen;
or a pharmaceutically acceptable salt thereof
C. Compounds according to formula 1, having the structure 
X is selected from NH, sulfur, or oxygen;
or a pharmaceutically acceptable salt thereof.
D. Compounds according to formula 1, having the structure 
wherein
R2 is hydrogen, amino, trifluoromethyl, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, cyano, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, phenylamino, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, dialkylamino of 2 to 12 carbon atoms, 
or a pharmaceutically acceptable salt thereof;
E. Compounds according to formula 1, having the structure 
R2 is hydrogen, amino, trifluoromethyl, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, cyano, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, phenylamino, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, dialkylamino of 2 to 12 carbon atoms, 
F. Compounds according to formula 1 having the structure 
R1 is hydrogen, hydroxymethyl, aminomethyl, N-alkylaminomethyl of 2-6 carbon atoms, N,N-dialkylaminomethyl of 3-12 carbon atoms, N-cycloalkylaminomethyl of 4-9 carbon atoms, N-cycloalkyl-N-alkylaminoalkyl of 5-16 carbon atoms, N,N-dicycloalkylaminomethyl of 7-18 carbon atoms, morpholino-N-methyl, piperidino-N-methyl, N-alkyl-piperazino-N-methyl wherein the alkyl group is 1-6 carbon atoms, azacycloalkyl-N-methyl of 3-6 carbon atoms, N-(hydroxyalkyl)aminomethyl of 3-7 carbon atoms, N,N-di(hydroxyalkyl)aminomethyl of 5-12 carbon atoms, N-(hydroxycycloalkyl)aminomethyl of 4-9 carbon atoms, N-(hydroxycycloalkyl)-N-(hydroxyalkyl)aminoalkyl of 6-16 carbon atoms, or N,N-di(hydroxycycloalkyl)aminomethyl of 7-18 carbon atoms;
R2 is hydrogen;
R3 is hydrogen, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, hydroxyalkyl of 2-6 carbon atoms; mercaptoalkyl of 2-6 carbon atoms, phenyl, benzyl, 
or a pharmaceutically acceptable salt thereof.
G. Compounds according to formula 1, having the structure 
R2 and R3 are hydrogen;
or a pharmaceutically acceptable salt thereof.
H. Compounds according to formula 1, having the structure 
R3 is hydrogen;
or a pharmaceutically acceptable salt thereof.
I. Compounds according to formula 1, having the structure 
R2 is hydrogen, amino, hydroxyamino, trifluoromethyl, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, alkenylsulfonamido of 2-6 carbon atoms, alkynylsulfonamido of 2-6 carbon atoms, cyano, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, N-alkyl-N-alkenylamino of 4 to 12 carbon atoms, N,N-dialkenylamino of 6-12 carbon atoms, phenylamino, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms, 
R3 is hydrogen, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, hydroxyalkyl of 2-6 carbon atoms; mercaptoalkyl of 2-6 carbon atoms, phenyl, benzyl, 
or a pharmaceutically acceptable salt thereof.
J. Compounds according to the formula 1, having the structure 
or a pharmaceutically acceptable salt thereof.
K. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R1 and R4 are hydrogen;
or a pharmaceutically acceptable salt thereof.
L. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R4 is hydrogen and one or two of the substituents R1, R2 and R3 are as defined above, the remaining being hydrogen;
or a pharmaceutically acceptable salt thereof.
M. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R4 is hydrogen and one or two of the substituents R1, R2 and R3 are as herein above described, the remaining being hydrogen;
or a pharmaceutically acceptable salt thereof.
N. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R2 is hydrogen, amino, trifluoromethyl, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, cyano, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, phenylamino, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, dialkylamino of 2 to 12 carbon atoms, 
or a pharmaceutically acceptable salt thereof.
O. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R2 is hydrogen, amino, trifluoromethyl, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, cyano, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, phenylamino, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, dialkylamino of 2 to 12 carbon atoms, 
or a pharmaceutically acceptable salt thereof.
P. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R1 is hydrogen, hydroxymethyl, aminomethyl, N-alkylaminomethyl of 2-6 carbon atoms, N,N-dialkylaminomethyl of 3-12 carbon atoms, N-cycloalkylaminomethyl of 4-9 carbon atoms, N-cycloalkyl-N-alkylaminoalkyl of 5-16 carbon atoms, N,N-dicycloalkylaminomethyl of 7-18 carbon atoms, morpholino-N-methyl, piperidino-N-methyl, N-alkyl-piperazino-N-methyl wherein the alkyl group is 1-6 carbon atoms, azacycloalkyl-N-methyl of 3-6 carbon atoms, N-(hydroxyalkyl)aminomethyl of 3-7 carbon atoms, N,N-di(hydroxyalkyl)aminomethyl of 5-12 carbon atoms, N-(hydroxycycloalkyl)aminomethyl of 4-9 carbon atoms, N-(hydroxycycloalkyl)-N-(hydroxyalkyl)aminoalkyl of 6-16 carbon atoms, or N,N-di(hydroxycycloalkyl)aminomethyl of 7-18 carbon atoms;
R2 is hydrogen;
R3 is hydrogen, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, hydroxyalkyl of 2-6 carbon atoms; mercaptoalkyl of 2-6 carbon atoms, phenyl, benzyl, 
or a pharmaceutically acceptable salt thereof.
Q. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R2 and R3 are hydrogen;
or a pharmaceutically acceptable salt thereof.
R. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R3 is hydrogen;
or a pharmaceutically acceptable salt thereof.
S. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R2 is hydrogen, amino, hydroxyamino, trifluoromethyl, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, alkenylsulfonamido of 2-6 carbon atoms, alkynylsulfonamido of 2-6 carbon atoms, cyano, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, N-alkyl-N-alkenylamino of 4 to 12 carbon atoms, N,N-dialkenylamino of 6-12 carbon atoms, phenylamino, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms, 
R3 is hydrogen, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, hydroxyalkyl of 2-6 carbon atoms; mercaptoalkyl of 2-6 carbon atoms, phenyl, benzyl, 
or a pharmaceutically acceptable salt thereof.
T. Compounds according to formula 1, having the structure 
Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
R1 and R4 are hydrogen;
or a pharmaceutically acceptable salt thereof.
Another group of preferred compounds of the present invention are those in which:
1) Ar is a phenyl ring which may be optionally mono-, di- or tri-substituted with a substituent selected from the group consisting of halogen, alkyl of 1-6 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, azido, hydroxyalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkanoyloxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, alkylthio of 1-6 carbon atoms, hydroxy, trifluoromethyl, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, benzoyl, amino, alkylamino of 1-6 carbon atoms, dialkylamino of 2-12 carbon atoms, alkanoylamino of 1-6 carbon atoms, alkenoylamino of 3-8 carbon atoms, alkynoylamino of 3-8 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, and benzoylamino; or
Ar is the radical: 
2) X is NH, S, or O;
3) R2 is hydrogen, amino, trifluoromethyl, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, cyano, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, phenylamino, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, alkanoyloxy of 1-6 carbon atoms, alkenoyloxy of 3-8 carbon atoms, alkynoyloxy of 3-8 carbon atoms, carbamoyl, N-alkylcarbamoyl of 2-7 carbon atoms, N,N-dialkylcarbamoyl of 3-13 carbon atoms, dialkylamino of 2 to 12 carbon atoms, 
It being especially preferred when R2 is H;
4) R1 is hydrogen, hydroxymethyl, aminomethyl, N-alkylaminomethyl of 2-6 carbon atoms, N,N-dialkylaminomethyl of 3-12 carbon atoms, N-cycloalkylaminomethyl of 4-9 carbon atoms, N-cycloalkyl-N-alkylaminoalkyl of 5-16 carbon atoms, N,N-dicycloalkylaminomethyl of 7-18 carbon atoms, morpholino-N-methyl, piperidino-N-methyl, N-alkyl-piperazino-N-methyl wherein the alkyl group is 1-6 carbon atoms, azacycloalkyl-N-methyl of 3-6 carbon atoms, N-(hydroxyalkyl)aminomethyl of 3-7 carbon atoms, N,N-di(hydroxyalkyl)aminomethyl of 5-12 carbon atoms, N-(hydroxycycloalkyl)aminomethyl of 4-9 carbon atoms, N-(hydroxycycloalkyl)-N-(hydroxyalkyl)aminoalkyl of 6-16 carbon atoms, or N,N-di(hydroxycycloalkyl)aminomethyl of 7-18 carbon atoms; and/or
5) R3 is hydrogen, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, hydroxyalkyl of 2-6 carbon atoms; mercaptoalkyl of 2-6 carbon atoms, phenyl, benzyl, 
it being especially preferred when R3 is hydrogen;
Specifically preferred compounds of this invention include:
a) 4-(4-phenoxyanilino)benzo[g]quinoline-3-carbonitrile,
b) 4-(3-chloro-4-fluoroanilino)benzo[g]quinoline-3-carbonitrile,
c) 4-(4-chloro-5-methoxy-2-methylanilino)benzo[g]quinoline-3-carbonitrile,
d) 7,8-dimethoxy-4-(4-phenoxyanilino)benzo[g]quinoline-3-carbonitrile,
e) 4-(4-chloro-5-methoxy-2-methylanilino)-7,8-dimethoxybenzo[g]quinoline-3-carbonitrile,
f) 4-(3-chloro-4-fluoroanilino)-7,8-dimethoxybenzo[g]quinoline-3-carbonitrile,
g) 4-(2,4-dichloroanilino)-7,8-dimethoxybenzo[g]quinoline-3-carbonitrile,
h) 4-(2,4-dichloroanilino)-7,8-dihydroxybenzo[g]quinoline-3-carbonitrile,
i) 8-(3,4,5-trimethoxyanilino)-3H-[1,2,3]triazolo[4,5-g]quinoline-7-carbonitrile,
j) 9-(4-chloro-5-methoxy-2-methylanilino)pyrido[2,3-g]quinoxaline-8-carbonitrile,
k) 8-(5-methoxy-2-methylanilino)-2-{[2-(4-morpholinyl)ethyl]amino}imidazo[4,5-g]quinoline-7-carbonitrile,
l) 2-{[2-(4-morpholinyl)ethyl]amino}-8-(3,4,5-trimethoxyanilino)imidazo[4,5-g]quinoline-7-carbonitrile,
m) 2-amino-8-(4-phenoxyanilino)imidazo[4,5-g]quinoline-7-carbontrile,
n) 8-(3-bromo-phenylamino)imidazo[4,5-g]quinoline-7-carbonitrile,
o) 8-(2-bromo-4-chlorophenylamino)imidazo[4,5-g]quinoline-7-carbonitrile,
p) 8-(2-bromo-4-chloro-5-methoxyphenylamino)imidazo[4,5-g]quinoline-7-carbonitrile,
q) 8-(2-chloro-5-methoxyphenylamino)imidazo[4,5-g]quinoline-7-carbonitrile,
r) 8-(3-hydroxy-4-methylphenylamino)imidazo[4,5-g]quinoline-7-carbonitrile,
s) 8-(3,4,5-trimethoxyanilino)imidazo[4,5-g]quinoline-7-carbonitrile,
t) 8-(4-phenoxyanilino)imidazo[4,5-g]quinoline-7-carbonitrile,
u) 2-(chloromethyl)-8-(3,4,5-trimethoxyanilino)imidazo[4,5-g]quinoline-7-carbonitrile,
v) 2-(4-morpholinylmethyl)-8-(3,4,5-trimethoxyanilino)imidazo[4,5-g]quinoline-7-carbonitrile,
w) 8-(4-chloro-5-methoxy-2-methylanilino)-3-[2-(4-morpholinyl)ethyl]-3H-imidazo[4,5-g]quinoline-7-carbonitrile,
x) 3-[2-(4-morpholinyl)ethyl]-8-(4-phenoxyanilino)-3H-imidazo[4,5-g]quinoline-7-carbonitrile,
y) 8-[(4-chloro-5-methoxy-2-methylphenyl)amino]-thiazolo[4,5-g]quinoline-7-carbonitrile,
z) 4-(3-bromophenylamino)benzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
aa) 4-(4-chloro-2-fluorophenylamino)benzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
bb) 4-(2,4-dichlorophenylamino)benzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
cc) 4-(2,4-dichloro-5-methoxyphenylamino)benzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
dd) 4-(4-phenoxyphenylamino)benzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
ee) 4-(3-hydroxy-4-methylphenylamino)benzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
ff) 4-(4-chloro-2-fluorophenoxy)benzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
gg) 4-(4-chloro-5-methoxy-2-methylphenylamino)-8-nitrobenzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
hh) 8-amino-4-(4-chloro-5-methoxy-2-methylanilino)[1]benzothieno[3,2-b]pyridine-3-carbonitrile,
ii) 4-(3-bromoanilino)-6-nitro[1]benzothieno[3,2-b]pyridine-3-carbonitrile,
jj) 6-amino-4-(3-bromoanilino)[1]benzothieno[3,2-b]pyridine-3-carbonitrile,
kk) 4-(3-bromophenylamino)benzo[4,5]furo[3,2-b]pyridine-3-carbonitrile,
ll) 4-(4-chloro-2-fluorophenylamino)benzo[4,5]furo[3,2-b]pyridine-3-carbonitrile,
mm) 4-(3-hydroxy-4-methylphenylamino)benzo[4,5]furo[3,2-b]pyridine-3-carbonitrile,
nn) 4-(4-phenoxyphenylamino)benzo[4,5]furo[3,2-b]pyridine-3-carbonitrile,
oo) 4-(4-chloro-2-fluorophenoxy)benzo[4,5]furo[3,2-b]pyridine-3-carbonitrile,
pp) 4-(2,4-dichloroanilino)-8-nitro[1]benzothieno[3,2-b]pyridine-3-carbonitrile,
qq) 4-(3-bromoanilino)-8-nitro[1]benzothieno[3,2-b]pyridine-3-carbonitrile,
rr) 8-amino-4-(3-bromoanilino)[1]benzothieno[3,2-b]pyridine-3-carbonitrile,
ss) N-[4-(3-bromoanilino)-3-cyano[1]benzothieno[3,2-b]pyridin-8-yl]acrylamide,
tt) N-[4-(3-bromoanilino)-3-cyano[1]benzothieno[3,2-b]pyridin-6-yl]acrylamide,
uu) 4-(2,4-dichloroanilino)-7-methoxybenzo[g]quinoline-3-carbonitrile,
vv) 4-(2,4-dichloroanilino)-8-methoxybenzo[g]quinoline-3-carbonitrile,
ww) 4-(2,4-dichloroanilino)-7-hydroxybenzo[g]quinoline-3-carbonitrile,
xx) 4-(2,4-dichloroanilino)-8-hydroxybenzo[g]quinoline-3-carbonitrile,
yy) 4-(2,4-dichloroanilino)-7-[2-(dimethylamino)ethoxy]benzo[g]quinoline-3-carbonitrile,
zz) 4-(4-chloro-5-methoxy-2-methylanilino)-7-methoxy-8-(chloroethoxy)benzo[g]quinoline-3-carbonitrile,
aaa) 4-(4-chloro-5-methoxy-2-methylanilino)-8-methoxy-7-(chloroethoxy)benzo[g]quinoline-3-carbonitrile,
bbb) 4-(4-chloro-5-methoxy-2-methylanilino)-7-methoxy-8-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
ccc) 4-(4-chloro-5-methoxy-2-methylanilino)-8-methoxy-7-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
ddd) 4-(2,4-dichloro-5-methoxyanilino)-7-methoxy-8-(chloroethoxy)benzo[g]quinoline-3-carbonitrile,
eee) 4-(2,4-dichloro-5-methoxyanilino)-8-methoxy-7-(chloroethoxy)benzo[g]quinoline-3-carbonitrile,
fff) 4-(2,4-dichloro-5-methoxyanilino)-8-methoxy-7-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
ggg) 4-(2,4-dichloro-5-methoxyanilino)-7-methoxy-8-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
hhh) 4-(2,4-dichloro-5-methoxyanilino)-8-methoxy-7-[2-(4-methyl-1-piperazinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
iii) 4-(2,4-dichloro-5-methoxyanilino)-7-methoxy-8-[2-(4-methyl-1-piperazinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
jjj) 4-(4-chloro-5-methoxy-2-methylanilino)-8-methoxy-7-[2-(4-methyl-1-piperazinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
kkk) 4-(4-chloro-5-methoxy-2-methylanilino)-7-methoxy-8-[2-(4-methyl-1-piperazinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
lll) 4-[3-Chloro-4-(1-methyl-1H-imidazol-2-ylsulfanyl)phenylamino]-8-hydroxy-7-methoxybenzo[g]quinoline-3-carbonitrile,
mmm) 8-(2-Chloroethoxy)-4-[3-chloro-4-(1-methyl-1H-imidazol-2-ylsulfanyl)phenylamino]-7-methoxybenzo[g]quinoline-3-carbonitrile,
nnn) 4-[3-Chloro-4-(1-methyl-1H-imidazol-2-ylsulfanyl)phenylamino]-7-methoxy-8-(2-morpholin-4-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
ooo) 4-[3-Chloro-4-(1-methyl-1H-imidazol-2-ylsulfanyl)phenylamino]-8-(3-chloropropoxy)-7-methoxybenzo[g]quinoline-3-carbonitrile,
ppp) 4-[3-Chloro-4-(1-methyl-1H-imidazol-2-ylsulfanyl)phenylamino]-7-methoxy-8-(3-morpholin-4-yl-propoxy)benzo[g]quinoline-3-carbonitrile,
qqq) 4-[3-Chloro-4-(1-methyl-1H-imidazol-2-ylsulfanyl)phenylamino]-7-methoxy-8-[2-(4-methylpiperazin-1-yl)ethoxy]-benzo[g]quinoline-3-carbonitrile,
rrr) 4-[3-Chloro-4-(1-methyl-1H-imidazol-2-ylsulfanyl)phenylamino]-7-methoxy-8-(2-[1,2,3]triazol-2-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
sss) 4-[3-Chloro-4-(1-methyl-1H-imidazol-2-ylsulfanyl)phenylamino]-7-methoxy-8-(2-[1,2,3]triazol-1-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
ttt) 4-(2,4-Dichloro-5-methoxyphenylamino)-8-hydroxy-7-methoxybenzo[g]quinoline-3-carbonitrile,
uuu) 8-(3-Chloropropoxy)-4-(2,4-dichloro-5-methoxyphenylamino)-7-methoxybenzo[g]quinoline-3-carbonitrile,
vvv) 4-(2,4-Dichloro-5-methoxyphenylamino)-7-methoxy-8-(3-morpholin-4-yl-propoxy)benzo[g]quinoline-3-carbonitrile,
www) 4-(2,4-Dichloro-5-methoxyphenylamino)-7-methoxy-8-(2-[1,2,3]triazol-2-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
xxx) 4-(2,4-Dichloro-5-methoxyphenylamino)-7-methoxy-8-(2-[1,2,3 ]triazol-1-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
yyy) 4-(2,4-Dichloro-5-methoxyanilino)-7,8-dimethoxybenzo[b][1,8]naphthyridine-3-carbonitrile,
zzz) 8-(2-Chloroethoxy)-4-(2,4-dichloro-5-methoxyanilino)-7-methoxybenzo [b][1,8]naphthyridine-3-carbonitrile,
aaaa) 4-(2,4-Dichloro-5-methoxyanilino)-7-methoxy-8-[2-(4-morpholinyl)ethoxy]benzo[b][1,8]naphthyridine-3-carbonitrile,
bbbb) 8-(2-Chloroethoxy)-4-{3-chloro-4-[(1-methyl-1H-imidazol-2-yl)sulfanyl]anilino}-7-methoxybenzo[b][1,8]naphthyridine-3-carbonitrile,
cccc) 4-(2,4-Dichloro-5-methoxyanilino)-7-methoxy-8-[2-(4-methyl-1-piperazinyl)ethoxy]benzo[b][1,8]naphthyridine-3-carbonitrile,
dddd) 4-{3-Chloro-4-[(1-methyl-1H-imidazol-2-yl)sulfanyl]anilino}-7-methoxy-8-[2-(4-morpholinyl)ethoxy]benzo[b][1,8]naphthyridine-3-carbonitrile,
eeee) 4-(2,4-Dichloroanilino)-7,8-dimethoxybenzo[b][1,8]naphthyridine-3-carbonitrile,
ffff) 8-(2-Chloroethoxy)-4-(4-chloro-5-methoxy-2-methylanilino)-7-ethoxybenzo[g]quinoline-3-carbonitrile,
gggg) 8-(2-Chloroethoxy)-4-(2-chloro-4-fluoro-5-methoxyanilino)-7-methoxybenzo[g]quinoline-3-carbonitrile,
hhhh) 7-(2-Chloroethoxy)-4-(2-chloro-4-fluoro-5-methoxyanilino)-8-methoxybenzo[g]quinoline-3-carbonitrile,
iiii) 8-(2-Chloroethoxy)-4-(2-chloro-5-methoxy-4-methylphenylamino)-7-methoxybenzo[g]quinoline-3-carbonitrile,
jjjj) 7-(2-Chloroethoxy)-4-(2-chloro-5-methoxy-4-methylphenylamino)-8-methoxybenzo[g]quinoline-3-carbonitrile,
kkkk) 7-(2-Chloroethoxy)-4-(3-chloro-4-fluoroanilino)-8-methoxybenzo[g]quinoline-3-carbonitrile,
llll) 8-(2-Chloroethoxy)-4-(3-chloro-4-fluoroanilino)-7-methoxybenzo[g]quinoline-3-carbonitrile,
mmmm) 4-(4-Benzyloxy-3-chlorophenylamino)-7-(2-chloroethoxy)-8-methoxybenzo[g]quinoline-3-carbonitrile,
nnnn) 4-(4-Benzyloxy-3-chlorophenylamino)-8-(2-chloroethoxy)-7-methoxybenzo[g]quinoline-3-carbonitrile,
oooo) 7-(2-Chloroethoxy)-4-(3-chloro-4-phenoxyphenylamino)-8-methoxybenzo[g]quinoline-3-carbonitrile,
pppp) 8-(2-Chloroethoxy)-4-(3-chloro-4-phenoxyphenylamino)-7-methoxybenzo[g]quinoline-3-carbonitrile,
qqqq) 4-(4-Chloro-5-methoxy-2-methylanilino)-8-ethoxy-7-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
rrrr) 4-(4-Chloro-5-methoxy-2-methylanilino)-7-ethoxy-8-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
ssss) ({2[4-(4-Chloro-5-methoxy-2-methylphenylamino)-3-cyano-8-ethoxybenzo[g]quinoline-7-yloxy]-ethyl}-ethoxycarbonylmethyl-amino)-acetic acid ethyl ester,
tttt) ({2-[4-(4-Chloro-5-methoxy-2-methylphenylamino)-3-cyano-7-ethoxybenzo[g]quinoline-8-yloxy]-ethyl}-ethoxycarbonylmethylamino)-acetic acid ethyl ester,
uuuu) 2-(Carbamoylmethyl-{2-[4-(4-chloro-5-methoxy-2-methylphenylamino)-3-cyano-7-ethoxybenzo[g]quinolin-8-yloxy]-ethyl}-amino)-acetamide,
vvvv) 4-(2,4-Dichloroanilino)-7-methoxy-8-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
wwww) 4-(2,4-Dichloroanilino)-8-methoxy-7-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
xxxx) 8-Methoxy-7-[2-(4-methyl-1-piperazinyl)ethoxy]-4-(3,4,5-trimethoxyanilino)benzo[g]quinoline-3-carbonitrile,
yyyy) 7-Methoxy-8-[2-(4-methyl-1-piperazinyl)ethoxy]-4-(3,4,5-trimethoxyanilino)benzo[g]quinoline-3-carbonitrile,
zzzz) 7-Methoxy-8-[2-(4-morpholinyl)ethoxy]-4-(3,4,5-trimethoxyanilino)benzo[g]quinoline-3-carbonitrile,
aaaaa) 8-Methoxy-7-[2-(4-morpholinyl)ethoxy]-4-(3,4,5-trimethoxyanilino)benzo[g]quinoline-3-carbonitrile,
bbbbb) 4-(2-Chloro-4-fluoro-5-methoxyanilino)-8-methoxy-7-[2-(4-methyl-1-piperazinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
ccccc) 4-(2-Chloro-5-methoxy-4-methylanilino)-8-methoxy-7-[2-(4-methyl-1-piperazinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
ddddd) 4-(2-Chloro-5-methoxy-4-methylanilino)-7-methoxy-8-[2-(4-methyl-1-piperazinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
eeeee) 4-(2,4-Dichloro-5-methoxyanilino)-7-[2-(4-hydroxy-1-piperidinyl)ethoxy]-8-methoxybenzo[g]quinoline-3-carbonitrile,
fffff) 4-(3-Chloro-4-fluoroanilino)-7-methoxy-8-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
ggggg) 4-(2,4-Dichloro-5-methoxyanilino)-8-[2-(4-hydroxy-1-piperidinyl)ethoxy]-7-methoxybenzo[g]quinoline-3-carbonitrile,
hhhhh) 4-(2-Chloro-5-methoxy-4-methylanilino)-8-methoxy-7-[2-(4-hydroxy-1-piperidinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
iiiii) 4-(2-Chloro-5-methoxy-4-methylanilino)-7-methoxy-8-[2-(4-hydroxy-1-piperidinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
jjjjj) 4-(2-Chloro-4-fluoro-5-methoxyanilino)-8-methoxy-7-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
kkkkk) 4-(2-Chloro-4-fluoro-5-methoxyanilino)-7-methoxy-8-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
lllll) 4-(2-Chloro-4-fluoro-5-methoxyanilino)-7-methoxy-8-[2-(4-methyl-1-piperazinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
mmmmm) 4-(3-Chloro-4-fluoroanilino)-8-methoxy-7-[2-(4-morpholinyl)ethoxy]benzo[g]quinoline-3-carbonitrile,
nnnnn) 4-(3-Chloro-4-phenoxyphenylamino)-7-methoxy-8-(2-morpholin-4-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
ooooo) 4-(3-Chloro-4-phenoxyphenylamino)-8-methoxy-7-(2-morpholin-4-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
ppppp) 4-(2-Chloro-5-methoxy-4-methylphenylamino)-8-methoxy-7-(2-morpholin-4-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
qqqqq) 4-(2-Chloro-5-methoxy-4-methylphenylamino)-7-methoxy-8-(2-morpholin-4-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
rrrrr) 4-(4-Benzyloxy-3-chlorophenylamino)-8-methoxy-7-(2-morpholin-4-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
sssss) 4-(4-Benzyloxy-3-chlorophenylamino)-7-methoxy-8-(2-morpholin-4-yl-ethoxy)benzo[g]quinoline-3-carbonitrile,
ttttt) 8-(Benzyloxy)-4-[(2-chloro-4-fluoro-5-methoxyphenyl)amino]-7-methoxybenzo[g]quinoline-3-carbonitrile, and
uuuuu) 4-[(2-Chloro-4-fluoro-5-methoxyphenyl)amino]-8-hydroxy-7-methoxybenzo[g]quinoline-3-carbonitrile,
or a pharmaceutically acceptable salt thereof.
Also included in the present invention are compounds useful as intermediates for producing the above compounds of formula 1. Such intermediates specifically include the following:
a) 4-oxo-1,4-dihydrobenzo[g]quinoline-3-carbonitrile,
b) 4-chlorobenzo[g]quinoline-3-carbonitrile,
c) 3-(dimethylaminomethyleneamino)-6,7-dimethoxynaphthalene-2-carboxylic acid methyl ester,
d) 7,8-dimethoxy-4-oxo-1,4-dihydrobenzo[g]quinoline-3-carbonitrile,
e) 4-chloro-7,8-dimethoxybenzo[g]quinoline-3-carbonitrile,
f) 7-chloro-6-nitro-4-oxo-1-{[2-(trimethylsilyl)ethoxy]methyl}-1,4-dihydro-3-quinolinecarbonitrile,
g) 6,7-diamino-4-oxo-1-(2-trimethylsilanyl-ethoxymethyl)-1,4-dihydro-quinoline-3-carbonitrile,
h) 8-oxo-5-{[2-(trimethylsilyl)ethoxy]methyl}-5,8-dihydro[1,2,3]triazolo[4,5-g]quinoline-7-carbonitrile,
i) 8-oxo-5,8-dihydro[1,2,3]triazolo[4,5-g]quinoline-7-carbonitrile,
j) 8-chloro[1,2,3]triazolo[4,5-g]quinoline-7-carbonitrile,
k) 2-amino-8-oxo-5-{[2-(trimethylsilyl)ethoxy]methyl-5,8-dihydroimidazo[4,5-g]quinoline-7-carbonitrile,
l) 2-amino-8-oxo-5,8-dihydroimidazo[4,5-g]quinoline-7-carbonitrile,
m) 2-amino-8-chloroimidazo[4,5-g]quinoline-7-carbonitrile,
n) 8-oxo-5,8-dihydroimidazo[4,5-g]quinoline-7-carbonitrile,
o) 8-chloroimidazo[4,5-g]quinoline-7-carbonitrile,
p) 7-cyanoimidazo[4,5-g]quinolin-8-yl(3,4,5-trimethoxyphenyl)formamide,
q) 7-cyanoimidazo[4,5-g]quinolin-8-yl(4-phenoxyphenyl)formamide,
r) 7-{[2-(4-morpholinyl)ethyl]amino}-6-nitro-4-oxo-1-{[2-(trimethylsilyl)ethoxy]methyl}-1,4-dihydro-3-quinolinecarbonitrile,
s) 6-amino-7-{[2-(4-morpholinyl)ethyl]amino}-4-oxo-1-{[2-(trimethylsilyl)ethoxy]methyl}-1,4-dihydro-3-quinolinecarbonitrile,
t) 3-[2-(4-morpholinyl)ethyl]-8-oxo-5,8-dihydro-3H-imidazo[4,5-g]quinoline-7-carbonitrile,
u) 8-chloro-3-[2-(4-morpholinyl)ethyl]-3H-imidazo[4,5-g]quinoline-7-carbonitrile,
v) 1,4-dihydro-7-mercapto-6-nitro-4-oxo-1-[[2-(trimethylsilyl)ethoxy]methyl]-3-quinolinecarbonitrile,
w) 8-hydroxy[1,3]thiazolo[4,5-g]quinoline-7-carbonitrile,
x) 3-(dimethylaminomethyleneamino)benzo[b]thiophene-2-carboxylic acid methyl ester,
y) 4-hydroxybenzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
z) 4-chlorobenzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
aa) 4-hydroxy-8-nitrobenzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
bb) 4-chloro-8-nitrobenzo[4,5]thieno[3,2-b]pyridine-3-carbonitrile,
cc) 4-chloro-6-nitro[1]benzothieno[3,2-b]pyridine-3-carbonitrile,
dd) 3-(dimethylaminomethyleneamino)benzofuran-2-carboxylic acid ethyl ester,
ee) 4-hydroxybenzo[4,5]furo[3,2-b]pyridine-3-carbonitrile,
ff) 4-chlorobenzo[4,5]furo[3,2-b]pyridine-3-carbonitrile,
gg) 7-methoxy-4-oxo-1,4-dihydrobenzo[g]quinoline-3-carbonitrile,
hh) 8-methoxy-4-oxo-1,4-dihydrobenzo[g]quinoline-3-carbonitrile,
ii) 4-chloro-7-methoxybenzo[g]quinoline-3-carbonitrile,
jj) 4-chloro-8-methoxybenzo[g]quinoline-3-carbonitrile,
kk) ethyl 7-(2-chloroethoxy)-6-methoxy-3-nitro-2-naphthoate,
ll) ethyl 6-(2-chloroethoxy)-7-methoxy-3-nitro-2-naphthoate,
mm) ethyl 3-amino-7-(2-chloroethoxy)-6-methoxy-2-naphthoate,
nn) ethyl 3-amino-6-(2-chloroethoxy)-7-methoxy-2-naphthoate,
oo) 8-(2-chloroethoxy)-7-methoxy-4-oxo-1,4-dihydrobenzo[g]quinoline-3-carbonitrile,
pp) 7-(2-chloroethoxy)-8-methoxy-4-oxo-1,4-dihydrobenzo[g]quinoline-3-carbonitrile,
qq) 4-chloro-7-methoxy-8-(2-chloroethoxy)benzo[g]quinoline-3-carbonitrile,
rr) 4-chloro-8-methoxy-7-(2-chloroethoxy)benzo[g]quinoline-3-carbonitrile,
ss) 7,8-dimethoxy-4-oxo-1,4-dihydrobenzo[b][1,8]naphthyridine-3-carbonitrile,
tt) 4-chloro-7,8-dimethoxybenzo[b][1,8]naphthyridine-3-carbonitrile,
uu) 8-(2-chloroethoxy)-7-methoxy-4-oxo-1,4-dihydrobenzo[b][1,8]naphthyridine-3-carbonitrile, and
vv) 4-chloro-8-(2-chloroethoxy)-7-methoxybenzo[b][1,8]naphthyridine-3-carbonitrile.
The compounds and intermediates of this invention encompassed by Formula 6 may be prepared as described below and in Flowsheet 1 wherein Ar, X and n are hereinbefore defined. R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 are each, independently, hydrogen, halogen, hydroxy, amino, hydroxyamino, trifluoromethyl, trifluoromethoxy, mercapto, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, alkynyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, alkenylsulfonamido of 2-6 carbon atoms, alkynylsulfonamido of 2-6 carbon atoms, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, N-alkylcarbamoyl, N,N-dialkylcarbamoyl, N-alkyl-N-alkenylamino of 4 to 12 carbon atoms, N,N-dialkenylamino of 6-12 carbon atoms, phenylamino, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, or dialkylamino of 2 to 12 carbon atoms;
R7xe2x80x94(C(R6)2)gxe2x80x94Vxe2x80x94,Phxe2x80x94(C(R62)qxe2x80x94Wxe2x80x94(C(R6)2)kxe2x80x94Vxe2x80x94.
where V, R6, R7, W, Ph, g, k and q are as hereinabove defined.
Reaction of 3-amino-2-naphthoic acids (Formula 2) with dimethylformamide dimethyl acetal, with or without a solvent, gives intermediates of Formula 3. The reaction of 3 with the lithium anion of acetonitrile prepared by using a base such as n-butyllithium or the like in an inert solvent gives 3-cyano-4-oxo-1,4-dihydrobenzo[g]quinolines 4 or the 3-cyano-4-hydroxybenzo[g]quinoline tautomers thereof. Heating 4, with or without solvent, with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the corresponding 4-chloro-3-cyanobenzo[g]quinolines. Condensation of 4-chloro-3-cyanobenzo[g]quinolines with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5, HXxe2x80x94(CH2)nxe2x80x94Ar, wherein Ar, X and n are as hereinbefore defined, give the 3-cyanobenzo[g]quinolines of Formula 6. The condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in an alcohol solvent, or by using transition metal catalysts such as tris(dibenzylideneacetone)dipalladium(0) or the like, together with ligands such as, but not limited to 2-dicyclohexylphosphino-2xe2x80x2-(N,N-dimethylamino)biphenyl, and potassium phosphate or the like in an inert solvent. In those cases where the substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or the R and S optically active forms, respectively. In cases where the substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. When Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with the substituted 4-chloro-3-cyanobenzo[g]quinolines. Suitable protecting groups include, but are not limited to tert-butoxycarbonyl (BOC), xcex2-trimethylsilylethanesulfonamide (SES), benzyloxycarbonyl (CBZ) and benzyl (Bn) protecting groups. The first protecting group listed above can be removed from the final products of Formula 6 by treatment with an acid such as trifluoroactic acid, the second protecting group with a fluoride salt, such as cesium fluoride or tetrabutylammonium fluoride. The latter two protecting groups can be removed by catalytic hydrogenation or sodium in ammonia. In those cases where the Ar contains hydroxyl groups, the hydroxyl groups may first have to be protected prior to final product formation. Suitable protecting groups include, but are not limited to, t-butyldimethylsilyl, tetrahydropyranyl, or benzyl protecting groups. The first two protecting groups listed above can be removed from the final products of Formula 6 by treatment with an acid such as acetic acid or hydrochloric acid while the latter protecting group can be removed by catalytic hydrogenation. The 3-amino-2-naphthoic acids of Formula 2 are commercially available or can be prepared by procedures known in the art from compounds detailed by the following references: Zhu, Z.; Drach, J. C.; Townsend, L. B. J. Org. Chem., 63, 977-983, (1998); Kienzle, F. Helv. Chim. Acta., 63, 2364-2369, (1980), Kobayashi, K.; Kanno, C.; Seko, S.; Suginome, H. J. Chem. Soc., Perkin Trans. 1., 3111-317, (1992), Levy, L. A. Synth. Commun., 13, 639-48 (1983) and Moder, K. P.; Leonard, N. J. J. Am. Chem. Soc., 104, 2613-24 (1982).
It will be recognized by those skilled in the art that the 4-hydroxy substituent of the benzoquinoline tautomer may be converted to a leaving group such as halogen, tosyl, mesyl, aryl- or alkyl-sulfonate, preferably trifluoromethanesulfonate and the like. 
Intermediate 3 can also be prepared as described below and in Flowsheet 1xe2x80x2.
The reaction of substituted naphtho[2,3-c]furan-1,3-dione compounds (McOmie, John F. W.; Perry, David H. Synthesis (1973), Issue 7, 416-417) with an alcohol such as methanol, with or without a base such as sodium hydride, provides substituted 3-(methoxycarbonyl)-2-naphthoic acids as a mixture of geometric isomers if R1xe2x80x2 differs from R4xe2x80x2 and/or R2xe2x80x2 differs from R3xe2x80x2. Treatment of the 3-(methoxycarbonyl)-2-naphthoic acids with diphenylphosphoryl azide and a base such as triethylamine in an inert solvent, followed by workup with an aqueous acetone solution or the like, provides the corresponding substituted methyl 3-amino-2-naphthoates, which when reacted with dimethylformamide dimethyl acetal, with or without a solvent, provides intermediates of Formula 3 (and the geometric isomer 3xe2x80x2 if R1xe2x80x2 differs from R4xe2x80x2 and/or R2xe2x80x2 differs from R3xe2x80x2). Separation of the geometric isomers can be carried out by silica gel chromatography or other purification methods at any step in the preparation of intermediate of Formula 3. The above-mentioned chemical transformations can be carried out separately on each isomer. If a mixture of geometric isomers of Formula 3 and 3xe2x80x2 is converted to compounds of Formula 6, a chromatographic separation can be carried out on the mixture of products of Formula 6 or any of the intermediates formed in this sequence. 
Intermediate 4 can also be prepared as described below and in Flowsheet 1xe2x80x2.
The reaction of phenyl compounds with electron-rich R2xe2x80x2 and R3xe2x80x2 substituents, such as alkoxy of 1 to 6 carbons or, for example, a haloalkoxy moiety of the formula R7xe2x80x94(C(R6)2)gxe2x80x94Vxe2x80x94 where R6 is hydrogen, R7 is a halogen, V is oxygen and g=2-6, a solution of formalin and hydrochloric acid provides substituted 1,2-bis(chloromethyl)benzene intermediates. Heating these substituted 1,2-bis(chloromethyl)benzene intermediates with sodium acetate in acetic acid provides the corresponding substituted 1,2-bis(acetyloxymethyl)benzene compounds which can be converted to the corresponding 1,2-bis(hydroxymethyl)benzene intermediates by reaction with a ammonia-saturated methanol or aqueous sodium hydroxide. Oxidation of the substituted 1,2-bis(hydroxymethyl)benzene intermediates by oxalyl chloride, dimethyl sulfoxide and triethylamine in an inert solvent such as methylene chloride provides the substituted phthalaldehyde intermediates. Reaction of the substituted phthalaldehyde intermediates with an excess of a 3-nitropropanoate ester such as ethyl 3-nitropropanoate (as described by Kienzle, F. Helv. Chim. Acta., 63, 2364-2369, (1980)), and sodium ethoxide in ethanol provides the corresponding ethyl 3-nitro-2-naphthoate intermediates as a mixture of geometric isomers if R2xe2x80x2 differs from R3xe2x80x2. Reduction of the substituted ethyl 3-nitro-2-naphthoate intermediates by catalytic hydrogenation over palladium-on-carbon or platinum-on-carbon in tetrahydrofuran provides the substituted ethyl 3-amino-2-naphthoate intermediates as a mixture of geometric isomers if R2xe2x80x2 differs from R3xe2x80x2. Reaction of the substituted ethyl 3-amino-2-naphthoate intermediates with dimethylformamide dimethyl acetal, with or without a solvent, followed by reaction with the lithium anion of acetonitrile prepared by using a base such as n-butyllithium or the like, in an inert solvent, gives 3-cyano-4-oxo-1,4-dihydrobenzo[g]quinolines 4 (and the geometric isomer 4xe2x80x2 if R2xe2x80x2 differs from R3xe2x80x2) or the 3-cyano-4-hydroxybenzo[g]quinoline tautomers thereof Separation of the geometric isomers can be carried out by silica gel chromatography or other purification methods at any step in the preparation of intermediate of Formula 4. The above-mentioned chemical transformations can be carried out separately on each isomer. If a mixture of geometric isomers of Formula 4 and 4xe2x80x2 is converted to compounds of Formula 6, a chromatographic separation can be carried out on the mixture of products of Formula 6 or any of the intermediates formed in this sequence. 
Intermediate 4 can also be prepared as described below and in Flowsheet 1xe2x80x2xe2x80x3.
Bicyclo[4.2.0]octa-1(6),2,4-triene-7-carbonitriles with R1xe2x80x2, R2xe2x80x2 and R3xe2x80x2 substituents being alkoxy of 1 to 6 carbon atoms, alkyl of 1 to 6 carbon atoms, or benzyloxy moiety of the formula Phxe2x80x94(C(R6)2)qxe2x80x94Wxe2x80x94(C(R6)2)kxe2x80x94Vxe2x80x94 where R6 is hydrogen, W is a bond, V is oxygen and k=0, q=1, can be synthesized in regioisomerically pure form by procedures known in the art as detailed by the following references: Kametani, T. et al J. Het. Chem, 11, 179, (1974), Kametani, T.; kondoh, H.; Tsubuki, M.; Honda, T. J. Chem. Soc Perkin Trans. 1, 5 (1990), Kametani, T.; Kato, Honda, T. Fukumoto, K. J. Chem. Soc Perkin 1, 2001 (1990), Kametani, T.; Kajiwara, M.; Takahashi, T.; Fukumoto, K. Tetrahedron, 31, 949 (1975) and Honda, T. Toya, T. Heterocycles, 33, 291 (1992). The reaction of the substituted bicyclo[4.2.0]octa-1(6),2,4-triene-7-carbonitriles with a base such as sodium (bistrimethylsilyl)amide or n-butyllithium at xe2x88x9278xc2x0 C. and the like provides the corresponding anion xcex1 to the cyano group which is then reacted with a suitable electrophile such as a substituted diphenyl disulfide PhSSPh (where Ph is as hereinabove defined) to provide substituted 7-phenylsulfanylbicyclo[4.2.0]octa-1,3,5-triene-7-carbonitriles after warming to room temperature. Reaction of these intermediates with the magnesium bromide salt of an ester such as, but not limited to t-butyl acetate at 0xc2x0 C. in an inert solvent such as ether or tetrahydrofuran and the like provides the corresponding substituted 3-amino-3-(7-phenylsulfanyl-bicyclo[4.2.0]octa-1,3,5-trien-7-yl)-acrylic acid tert-butyl esters. Refluxing these adducts in a high boiling solvent such as dichlorobenzene or the like for 0.5 to 3 hours provides the substituted 3-amino-naphthalene-2-carboxylic acid tert-butyl esters. Reaction of the substituted 3-amino-2-naphthoate tert-butyl ester intermediates with dimethylfornamide dimethyl acetal, with or without a solvent, followed by reaction with the lithium anion of acetonitrile prepared by using a base such as n-butyllithium or the like, in an inert solvent, gives 3-cyano-4-oxo-1,4-dihydrobenzo[g]quinolines 4 or the 3-cyano-4-hydroxybenzo[g]quinoline tautomers thereof. 
Converting the R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 groups to R1, R2, R3 and R4 groups can be accomplished through any conventionally known techniques, for example:
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a methoxy group, it can be converted to the corresponding hydroxy group by reaction with a demethylating agent such as boron tribromide in an inert solvent or by heating with pyridinium chloride with or without solvent;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 is a benzyloxy group of the formula Phxe2x80x94(C(R6)2)qxe2x80x94Wxe2x80x94(C(R6)2)kxe2x80x94Vxe2x80x94 where R6 is hydrogen, W is a bond, V is oxygen and k=0, q=1, it can be converted to the corresponding hydroxy group by reaction with a debenzylating agent such as boron tribromide in an inert solvent, trifluoroacetic acid or catalytic hydrogenation with a catalyst such as palladium-on-carbon;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a hydroxy group, it can be converted to the corresponding alkanoyloxy group of 1-6 carbon atoms by reaction with an appropriate carboxylic acid chloride, anhydride, or mixed anhydride in a inert solvent using pyridine or a trialkylamine as a base;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a hydroxy group, it can be converted to the corresponding alkenoyloxy group of 1-6 carbon atoms by reaction with an appropriate carboxylic acid chloride, anhydride, or mixed anhydride in an inert solvent using pyridine or a trialkylamine as a base;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a hydroxy group, it can be converted to the corresponding alkynoyloxy group of 1-6 carbon atoms by reaction with an appropriate carboxylic acid chloride, anhydride, or mixed anhydride in a inert solvent using pyridine or a trialkylamine as a base;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a hydroxy group, it can be converted to the corresponding groups: 
wherein R5 is as defined hereinabove, by the reaction in an inert solvent with an alkyl or phenyl substituted isocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90O, or using a base such as pyridine, with a reagent (R5)2NCOCl;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a hydroxy group, it can be converted to the corresponding groups: 
wherein V is oxygen, R6, R7, R8, R9, M, W, Het, Ph, p and q are as defined hereinabove and g=2-6 and k=2-4 by reacting with an appropriately substituted alcohol using triphenyl phosphine and diethyl azodicarboxylate in an inert solvent, or alternatively by first reacting with a reagent such as, but not limited to, a bromoalkyl chloride or chloroalkyl tosylate to provide an intermediate haloalkoxy group which can be converted to the above described groups by subsequent reaction with an appropriately substituted nucleophile;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a hydroxy group, it can be converted to a alkoxycarbonyl group of 2-7 carbon atoms by first converting to a trifluoromethanesulfonate using trifluoromethanesulfonate anhydride or N-phenyltrifluoromethylsulfonamide and a base such as triethylamine in an inert solvent, then reacting with carbon monoxide in an alcoholic solvent of 1-6 carbons in the presence of a palladium (0) catalyst such as palladium tetrakis triphenylphosphine;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is an alkoxycarbonyl group of 2-7 carbon atoms, it can be converted to the corresponding hydroxymethyl group by reduction with an appropriate reducing agent such as lithium borohydride, or lithium aluminum hydride in a inert solvent; the hydroxymethyl group, in turn, can be converted to the corresponding halomethyl group by reaction in an inert solvent with a halogenating reagent such as phosphorous tribromide to give a bromomethyl group, or phosphorus pentachloride to give a chloromethyl group. The hydroxymethyl group can be acylated with an appropriate acid chloride, anhydride, or mixed anhydride in an inert solvent using pyridine or a trialkylamine as a base to give the compounds of this invention with the corresponding alkanoyloxymethyl group of 2-7 carbon atoms, alkenoyloxymethyl group of 2-7 carbon atoms, or alkynoyloxymethyl group of 2-7 carbon atoms;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a halomethyl group, it can be converted to the corresponding groups: 
wherein R6, R7, R8, R9, M, W, Het, p and q are as defined hereinabove by reacting with the appropriately substituted alcohol, amine or mercaptan in an inert solvent such as dioxane or acetonitrile and a base such as triethylamine or potassium carbonate;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a alkoxycarbonyl group of 2-7 carbon atoms, it can be converted to the corresponding carboxy group by reaction with a strong base such as aqueous sodium hydroxide in an alcoholic solvent such as ethanol;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a carboxy group, it can be converted to a carbamoyl, N-alkylcarbamoyl or N,N-dialkylcarbamoyl of 4-12 carbon atoms by reaction in an inert solvent with a halogenating agent such as phosphorus oxychloride or oxalyl chloride, or alternatively activating by reaction with a coupling agent such as, but not limited to carbonyl diimidazole in an inert solvent such as dimethylformamide, followed by reaction with the appropriate amine;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is a carboxy group, it can be converted to an amino group by heating with diphenyl phosphoryl azide and t-butanol in an inert solvent such as dioxane, followed by treatment with a strong acid such as hydrochloric or trifluoroacetic acid;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is an amino group, it can be converted to the corresponding dialkylamino group of 2 to 12 carbon atoms by alkylation with at least two equivalents of an alkyl halide of 1 to 6 carbon atoms by heating in an inert solvent;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is an amino group, it can be converted to the corresponding alkylsulfonamido, alkenylsulfonamido, or alkynylsulfonamido group of 2 to 6 carbon atoms by the reaction with an alkylsulfonyl chloride, alkenylsulfonyl chloride, or alkynylsulfonyl chloride, respectively, in an inert solvent using a base such as triethylamine or pyridine;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is an amino group, it can be converted to the corresponding alkylamino group of 1 to 6 carbon atoms by alkylation with one equivalent of an alkyl halide of 1 to 6 carbon atoms by heating in an inert solvent or by reductive alkylation using an aldehyde of 1 to 6 carbon atoms and a reducing agent such as sodium cyanoborohydride in a protic solvent such as water or alcohol, or mixtures thereof;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is an amino group, it can be converted to the corresponding groups: 
wherein R5 is as defined hereinabove by reacting with the appropriately substituted carboxylic acid chloride or sulfonyl chloride or mixed anhydride (which is prepared from the corresponding carboxylic acid) in an inert solvent such as tetrahydrofuran (THF) in the presence of an organic base such as pyridine, triethylamine or N-methyl morpholine;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is an amino group, it can be converted to the corresponding groups: 
wherein R5 is as defined hereinabove, by the reaction in an inert solvent with an alkyl or phenyl substituted isocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90O, or an alkyl or phenyl substituted isothiocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90S;
where one or more of R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 of Formula 6 or an intermediate is an amino group, it can be converted to the corresponding groups: 
wherein R5 is as defined hereinabove, by reacting with phosgene in an inert solvent such as toluene in the presence of a base such as pyridine to give an isocyanate which, in turn, is treated with an excess of the alcohol R5xe2x80x94OH.
In those cases when the R1xe2x80x2, R2xe2x80x2, R3xe2x80x2 and R4xe2x80x2 substituents of Formula 6 or an intermediate may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the substituents may contain more than one asymmetric carbon atom, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases where the R1xe2x80x2, R2xe2x80x2, R3xe2x80x2, R4xe2x80x2, R5, R6, R7, R8, R9 and Het substituents of Formula 6 or an intermediate contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups during the reaction sequence. The same amine or alcohol protecting groups described hereinabove can be used and they can be removed from the products of Formula 6 as previously described.
The preparation of the compounds and intermediates of this invention encompassed by Formula 13 is described below and in Flowsheet 2 where Ar, X and n are as hereinabove defined.
According to the sequence of reaction outlined in Flowsheet 2, a quinoline-3-carboxylic acid ester of Formula 7 is hydrolyzed with base to furnish a carboxylic acid of Formula 8. The carboxylic acid group of 8 is converted to an acyl imidazole by heating it with carbonyldiimidazole in an inert solvent such as dimethylformamide (DMF) followed by the addition of ammonia to give the amide 9. Dehydration of the amide functional group with the dehydrating agent, cyanuric chloride in dimethylformamide (DMF), gives the 3-cyano-4-quinolone of Formula 10. Deprotonation of 10 with sodium hydride in anhydrous dimethylformamide (DMF), followed by reaction with 2-(trimethylsilyl)ethoxymethyl (SEM) chloride provides a 4-quinolone of Formula 11. By heating 11 with sodium azide in dimethylsulfoxide (DMSO), it can be converted to an azide, which is reduced to the diamine of Formula 12 by catalytic hydrogenation over palladium-on-carbon or platinum-on-carbon in tetrahydrofuran. Reaction of 12 with nitrous acid provides 13. Refluxing 13 in formic acid provides the 7-cyano-8-oxo-5,8-dihydrotriazolo[4,5-g]quinoline 14 or the 7-cyano-8-hydroxytriazolo[4,5-g]quinoline tautomer thereof Heating 14 with or without solvent with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the corresponding 7-cyano-8-chlorotriazolo[4,5-g]quinoline. Condensation of 7-cyano-8-chlorotriazolo[4,5-g]quinoline with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5 gives the 7-cyano-triazolo[4,5-g]quinolines of Formula 15; this condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in alcohol solvents, and the like. In those cases where the Ar substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 5 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with 7-cyano-8-chlorotriazolo[4,5-g]quinoline. The same amine or alcohol protecting groups described hereinabove can be used and they can be removed from the products 15 as previously described.
The quinoline-3-carboxylic acid ester of Formula 7 needed to prepare the compounds of this invention are either already known in the art or can be prepared by procedures known in the art as detailed in the following reference: Koga, Hiroshi; Itoh, Akira; Murayama, Satoshi; Suzue, Seigo; Irikura, Tsutomu, J. Med. Chem., 23, 1358 (1980). 
The preparation of the compounds and intermediates of this invention encompassed by Formula 19 is described below and in Flowsheet 3 where Ar, X and n are as hereinabove defined.
Heating 10 with or without solvent with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the intermediate 4,7-dichloro-6-nitro-3-quinolinecarbonitrile. Condensation of 4,7-dichloro-6-nitro-3-quinolinecarbonitrile with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5 gives the 7-cyano-triazolo[4,5-g]quinolines of Formula 16; this condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in an alcohol solvents, and the like. Heating 16 with sodium azide in dimethylsulfoxide (DMSO), provides the corresponding azides, which are reduced to the diamines of Formula 17 by catalytic hydrogenation over palladium-on-carbon or platinum-on-carbon in tetrahydrofuran. Reaction of 17 with 2,3-dihydroxy-1,4-dioxane of Formula 18 in an inert solvent such as methanol provides the pyrido[2,3-g]quinoxaline-8-carbonitriles of Formula 19. In those cases where the Ar substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 5 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with 4,7-dichloro-6-nitro-3-quinoline carbonitrile. The same amine or alcohol protecting groups hereinabove can be used and they can be removed from the products 19 as previously described. 
The preparation of the compounds and intermediates of this invention encompassed by Formula 23 is described below and in Flowsheet 4 where Ar, X and n are as hereinabove defined. G is selected from the group consisting of: alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, hydroxyalkyl of 2-6 carbon atoms; mercaptoalkyl of 2-6 carbon atoms, phenyl, benzyl, 
where R6, R7, R8, R9, M, W, Het, Ph, p and q are as defined hereinabove, g=2-6 and k=2-4.
Reaction of 17 with an isothiocyanate 20 provides a mixture of thioureas of Formulas 21 and 22. Heating the mixture of Formulas 21 and 22 with mercury (II) oxide and a catalytic amount of sulfur in an inert solvent such as dioxane provides the corresponding substituted 2-amino-7-cyanoimidazo[4,5-g]quinolines of Formula 23. In those cases where the Ar and/or G substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar and/or G substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 20 where G contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with 17. The same amine or alcohol protecting groups defined hereinabove can be used and they can be removed from the products 23 as previously described. 
The preparation of the compounds and intermediates of this invention encompassed by Formula 26 is described below and in Flowsheet 5 where Ar, X and n are as hereinabove defined.
Reaction of 12 with cyanogen bromide in an inert solvent such as methanol provides a compound of Formula 24. Refluxing 24 in formic acid with 4 equivalents of imidazole provides a compound of formula 25. Heating 25 with or without solvent with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the corresponding 2-amino-8-chloroimidazo[4,5-g]quinoline-7-carbonitrile. Condensation of 2-amino-8-chloroimidazo[4,5-g]quinoline-7-carbonitrile with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5 gives the 7-cyano imidazo[4,5-g]quinolines of Formula 26; this condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in an alcohol solvents, and the like. In those cases where the Ar substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 5 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with 2-amino-8-chloro-imidazo[4,5-g]quinoline-7-carbonitrile. The same amine or alcohol protecting groups described hereinabove can be used and they can be removed from the products 26 as previously described. 
The preparation of the compounds and intermediates of this invention encompassed by Formula 28 is described below and in Flowsheet 6 where Ar, X and n are as hereinabove defined.
Refluxing 12 in formic acid with 4 equivalents of imidazole provides a compound of formula 27. Heating 27 with or without solvent with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the corresponding 8-chloroimidazo[4,5-g]quinoline-7-carbonitrile. Condensation of 8-chloroimidazo[4,5-g]quinoline-7-carbonitrile with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5 gives the 7-cyano-imidazo[4,5-g]quinolines of Formula 28; this condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in an alcohol solvents, and the like. In those cases where the Ar substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 5 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with 8-chloroimidazo[4,5-g]quinoline-7-carbonitrile. The same amine or alcohol protecting groups described hereinabove can be used and they can be removed from the products 28 as previously described. 
An alternative preparation of the compounds and intermediates of this invention encompassed by Formula 28 is described below and in Flowsheet 7 where Ar, X and n are as hereinabove defined.
Refluxing intermediates of Formula 17 in diethoxymethyl acetate provides the 7-cyano-imidazo[4,5-g]quinolines of Formula 28 when X is oxygen or sulfur. When X is nitrogen with a hydrogen substituent, the corresponding 7-cyanoimidazo[4,5-g]quinolin-8-ylformamides are formed. Heating the 7-cyanoimidazo[4,5-g]quinolin-8-ylformamides with potassium carbonate in a solvent such as methanol or ethanol provides the compounds of Formula 28. In those cases where the Ar substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 17 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with diethoxymethyl acetate. The same amine or alcohol protecting groups described hereinabove can be used and they can be removed from the products 28 as previously described. 
The preparation of the compounds and intermediates of this invention encompassed by Formula 32 is described below and in Flowsheet 8 where Ar, X and n are as hereinabove defined; and
Gxe2x80x2 is selected from the group consisting of: hydrogen, alkyl of 1-6 carbon atoms, trifluoromethyl, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, thiol, hydroxyalkyl of 1-6 carbon atoms; mercaptoalkyl of 1-6 carbon atoms; halomethyl, alkoxycarbonyl of 2-7 carbon atoms, phenyl, benzyl, phenoxy;
R7xe2x80x94(C(R6)2)gxe2x80x94Vxe2x80x94, Phxe2x80x94(C(R6)2)qxe2x80x94Wxe2x80x94(C(R6)2)kxe2x80x94Vxe2x80x94.
where g, k, q, R6, R7, V, W and Ph are as defined hereinabove.
Reaction of 17 with a carboxylic acid chloride of Formula 29 with a base such as pyridine, diethylaniline or triethylamine with or without an inert solvent such as tetrahydrofuran (THF) provides mixtures of compounds of Formula 30 and 31. Heating the mixture of Formulas 30 and 31 in formic acid or acetic acid provides the corresponding substituted 7-cyano-imidazo[4,5-g]quinolines of Formula 32. Alternatively, intermediates 17 can be directly converted to substituted 7-cyano-imidazo[4,5-g]quinolines of Formula 32 by reaction with Gxe2x80x2-C(Lxe2x80x2)3, where Lxe2x80x2 is chloro, hydroxy, alkoxy, alkylthio, phenoxy, thiophenoxy or dimethylamine, or two Lxe2x80x2 groups can be taken together to form xe2x95x90S, xe2x95x90NH, xe2x95x90O or xe2x95x90Se substituents, using acidic conditions (Hagen, H; Kohler, R.-D.; Fleig, H. Liebigs Ann. Chem., 1216 (1980), or basic reaction conditions (Webb, R. L. et al, J. Heterocycl. Chem., 24, 275 (1987), McKee, R. L.; Mckee, M. K.; Bost, R. W. J. Am. Chem. Soc., 68, 1904 (1946), Allen, J. A.; Deacon, B. D. Org. Synth., 30,56 (1950)) or by using a strongly dehydrating solvent such as polyphosphoric acid (Hein, D. W.; Leavitt, J. J. J. Am. Chem. Soc., 79, 427 (1957), or by using agents such as 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (Corrol, F. I.; Coleman, M. C. J. Med. Chem., 18, 318 (1975)) or by heating in an inert solvent (Cohen, V. I.; Pourabass, J. Heterocycl. Chem., 14, 1321 (1977)). In those cases, in intermediates 29 or Gxe2x80x2-C(Lxe2x80x2)3 where Gxe2x80x2 contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with 17. The same amine or alcohol protecting groups as defined hereinabove can be used and they can be removed from the products 32 as previously described. In those cases where the Ar and/or G1 substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. 
Converting the Gxe2x80x2 groups of Formula 32 to R2 groups can be accomplished through any conventionally known techniques.
The preparation of the compounds and intermediates of this invention encompassed by Formula 36 is described below and in Flowsheet 9 where Ar, X, Gxe2x80x2 and n are as hereinabove defined; and
Gxe2x80x3 is hydrogen, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, hydroxyalkyl of 2-6 carbon atoms; mercaptoalkyl of 2-6 carbon atoms, phenyl, benzyl, 
where R6, R7, R8, R9, M, W, Het, Ph, p and q are as hereinabove defined and g=2-6 and k=2-4.
By heating 11 with amines of Formula 33 in an inert solvent such as acetonitrile or dimethyl sulfoxide (DMSO), followed by catalytic hydrogenation over palladium on carbon in tetrahydrofuran and ethanol, it can be converted to compounds of Formula 34. The compounds of Formula 34 can be converted to compounds of Formula 35 by reaction with Gxe2x80x2xe2x80x94C(Lxe2x80x2)3, where Lxe2x80x2 is chloro, hydroxy, alkoxy, alkylthio, phenoxy, thiophenoxy or dimethylamine, or two Lxe2x80x2 groups can be taken together to form xe2x95x90S, xe2x95x90NH, xe2x95x90O or xe2x95x90Se substituents, using acidic conditions (Hagen, H; Kohler, R.-D.; Fleig, H. Liebigs Ann. Chem., 1216 (1980), or basic reaction conditions (Webb, R. L. et al, J. Heterocycl. Chem., 24, 275 (1987), McKee, R. L.; Mckee, M. K.; Bost, R. W. J. Am. Chem. Soc., 68, 1904 (1946), Allen, J. A.; Deacon, B. D. Org. Synth., 30, 56 (1950)) or by using a strongly dehydrating solvent such as polyphosphoric acid (Hein, D. W.; Leavitt, J. J. J. Am. Chem. Soc., 79, 427 (1957), or by using agents such as 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (Corrol, F. I.; Coleman, M. C. J. Med. Chem., 18, 318 (1975)) or by heating in an inert solvent (Cohen, V. I.; Pourabass, J. Heterocycl. Chem., 14, 1321 (1977)). Heating 35 with or without solvent with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the corresponding 8-chloroimidazo[4,5-g]quinoline-7-carbonitriles. Condensation of 8-chloroimidazo[4,5-g]quinoline-7-carbonitriles with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5 gives the 7-cyano-imidazo[4,5-g]quinolines of Formula 36; this condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in an alcohol solvent, and the like. In those cases where the Ar and/or Gxe2x80x2 and/or Gxe2x80x3 substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar and/or Gxe2x80x2 and/or Gxe2x80x3 substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 33 where Gxe2x80x3 contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with 11. In those cases, in intermediates 5 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with 8-chloroimidazo[4,5-g]quinoline-7-carbonitriles. The same amine or alcohol protecting groups described hereinabove can be used and they can be removed from the products 36 as previously described. 
Converting the Gxe2x80x2 groups of Formula 36 or Formula 35 to R2 groups and the Gxe2x80x3 groups of Formula 36 or Formula 35 to R3 groups can be accomplished through any conventionally known techniques.
The preparation of the compounds and intermediates of this invention encompassed by Formula 39 is described below and in Flowsheet 10 where Ar, X, Gxe2x80x2 and n are as hereinabove defined.
By heating 11 with sodium sulfide in dimethylsulfoxide (DMSO), followed by catalytic hydrogenation over palladium on carbon in tetrahydrofuran and ethanol, it can be converted to a compound of Formula 37. Refluxing 37 in formic acid with 4 equivalents of imidazole provides a compound of formula 38. The compounds of Formula 37 can be converted to compounds of Formula 38 by reaction with Gxe2x80x2xe2x80x94C(Lxe2x80x2)3, where Lxe2x80x2 is chloro, hydroxy, alkoxy, alkylthio, phenoxy, thiophenoxy or dimethylamine, or two Lxe2x80x2 groups can be taken together to form xe2x95x90S, xe2x95x90NH, xe2x95x90O or xe2x95x90Se substituents, using acidic conditions (Hagen, H; Kohler, R.-D.; Fleig, H. Liebigs Ann. Chem., 1216 (1980), or basic reaction conditions (Tawins, A.; Hsiu, R. K.-C. Can. J. Chem., 49, 4054 (1971)) or by using a strongly dehydrating solvent such as polyphosphoric acid (Hein, D. W.; Leavitt, J. J. J. Am. Chem. Soc., 79, 427 (1957), or by using agents such as phosphorus oxychloride (Davis, C. S. J. Pharm. Sci., 51, 1111 (1962)) or by heating in an inert solvent (Campaigne, E.; Van Verth, J. E. J. Org. Chem., 23, 1344 (1958), George, B.; Papadopoulos, E. P. J. Org. Chem., 42, 2530 (1977). Heating 38 with or without solvent with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the corresponding 7-cyano-8-chlorothiazolo[4,5-g]quinoline. Condensation of a substituted 7-cyano-8-chlorothiazolo[4,5-g]quinoline with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5 gives the 7-cyanothiazolo[4,5-g]quinolines of Formula 39; this condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in an alcohol solvent, and the like. In those cases where Ar and/or Gxe2x80x2 may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 5 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with 7-cyano-8-chlorothiazolo[4,5-g]quinoline. The same amine or alcohol protecting groups described hereinabove can be used and they can be removed from the products 39 as previously described. 
Converting the Gxe2x80x2 groups of Formula 39 to R2 groups can be accomplished through any conventionally known techniques.
The preparation of the compounds and intermediates of this invention encompassed by Formula 44 is described below and in Flowsheet 11 where Ar, X and n are as hereinabove defined.
Q1, Q2, Q3 and Q4 are each, independently, hydrogen, halogen, hydroxy, amino, hydroxyamino, trifluoromethyl, trifluoromethoxy, mercapto, alkyl of 1-6 carbon atoms, cycloalkyl of 3-8 carbon atoms, alkenyl of 2-6 carbon atoms, alkynyl of 2-6 carbon atoms, alkenyloxy of 2-6 carbon atoms, alkynyloxy of 2-6 carbon atoms, hydroxyalkyl of 1-6 carbon atoms, mercaptoalkyl of 1-6 carbon atoms, halomethyl, alkoxymethyl of 2-7 carbon atoms, alkoxy of 1-6 carbon atoms, cycloalkoxy of 3-8 carbon atoms, alkylthio of 1-6 carbon atoms, cycloalkylthio of 3-8 carbon atoms, alkylsulphinyl of 1-6 carbon atoms, alkylsulfonyl of 1-6 carbon atoms, alkylsulfonamido of 1-6 carbon atoms, alkenylsulfonamido of 2-6 carbon atoms, alkynylsulfonamido of 2-6 carbon atoms, cyano, nitro, carboxy, alkoxycarbonyl of 2-7 carbon atoms, alkanoyl of 2-7 carbon atoms, N-alkylcarbamoyl, N,N-dialkylcarbamoyl, N-alkyl-N-alkenylamino of 4 to 12 carbon atoms, N,N-dialkenylamino of 6-12 carbon atoms, phenylamino, benzylamino, phenoxy, phenyl, thiophenoxy, benzyl, alkylamino of 1-6 carbon atoms, dialkylamino of 2 to 12 carbon atoms, 
where V, R6, R7, R8, R9, M, W, Het, Ph, g, k, p and q are as hereinabove defined.
By reacting substituted 2-nitrobenzonitriles of Formula 40 with methyl thioglycolate and a base such as potassium hydroxide or triethylamine in an inert solvent such as dimethyl sulfoxide (DMSO) or aqueous dimethyl formamide (DMF) with or without heating provides compounds of Formula 41. Heating the substituted aniline of Formula 41 with dimethylformamide dimethyl acetal with or without a solvent gives intermediates of Formula 42. The reaction of 42 with from one to ten equivalents of acetonitrile using a base such as n-butyllithium, sodium methoxide or the like in an inert solvent gives the 4-oxo-1,4-dihydro[1]benzothieno[3,2-b]pyridine-3-carbonitrile 43, or the 4-hydroxy[1]benzothieno[3,2-b]pyridine-3-carbonitriles tautomers thereof. Heating 43 with or without solvent with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the corresponding 4-chloro[1]benzothieno[3,2-b]pyridine-3-carbonitriles. Condensation of 4-chloro[1]benzothieno[3,2-b]pyridine-3-carbonitriles with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5 gives the benzothieno[3,2-b]pyridine-3-carbonitriles of Formula 44; this condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in an alcohol solvents, and the like. In those cases where the Ar and/or Q1, Q2, Q3 and Q4 substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar and/or Q1, Q2, Q3 and Q4 substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 40 where Q1, Q2, Q3 and Q4 contain primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with methyl thioglycolate. In those cases, in intermediates 5 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with the 4-chloro[1]benzothieno[3,2-b]pyridine-3-carbonitriles. The same amine or alcohol protecting groups described hereinabove can be used and they can be removed from the products 44 as previously described.
The 2-nitrobenzonitriles of Formula 40 are either commercially available, or are already known to the art or can be prepared by procedures known in the art. 
Converting the Q1, Q2, Q3 and Q4 groups to R1, R2, R3 and R4 groups can be accomplished through any conventionally known techniques, for example:
Where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is a nitro group, it can be converted to the corresponding amino group by reduction using a reducing agent such as iron in acetic acid;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is an amino group, it can be converted to the corresponding dialkylamino group of 2 to 12 carbon atoms by alkylation with at least two equivalents of an alkyl halide of 1 to 6 carbon atoms by heating in an inert solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is a methoxy group, it can be converted to the corresponding hydroxy group by reaction with a demethylating agent such as boron tribromide in an inert solvent or by heating with pyridinium chloride with or without solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is an amino group, it can be converted to the corresponding alkylsulfonamido, alkenylsulfonamido, or alkynylsulfonamido group of 2 to 6 carbon atoms by the reaction with an alkylsulfonyl chloride, alkenylsulfonyl chloride, or alkynylsulfonyl chloride, respectively, in an inert solvent using a base such as triethylamine or pyridine;
where two of Q1, Q2, Q3 or Q4 of Formula 44 are contiguous methoxy groups, the corresponding compound with contiguous hydroxy groups can be prepared by using a demethylating agent such as boron tribromide in an inert solvent or by heating with pyridinium chloride with or without solvent.
Where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is an amino group, it can be converted with or without heating to the corresponding alkylamino group of 1 to 6 carbon atoms by alkylation with one equivalent of an alkyl halide of 1 to 6 carbon atoms or by reductive alkylation using an aldehyde of 1 to 6 carbon atoms and a reducing agent such as sodium cyanoborohydride in a protic solvent such as water or alcohol, or mixtures thereof;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is hydroxy, it can be converted to the corresponding alkanoyloxy, group of 1-6 carbon atoms by reaction with an appropriate carboxylic acid chloride, anhydride, or mixed anhydride in a inert solvent using pyridine or a trialkylamine as a base;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is hydroxy, it can be converted to the corresponding alkenoyloxy group of 1-6 carbon atoms by reaction with an appropriate carboxylic acid chloride, anhydride, or mixed anhydride in an inert solvent using pyridine or a trialkylamine as a base;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is hydroxy, it can be converted to the corresponding groups: 
wherein V is oxygen, R6, R7, R8, R9, M, W, Het, Ph, p and q are as hereinabove defined and g=2-6 and k=2-4 by reacting with the appropriately substituted alcohol using triphenyl phosphine and diethyl azodicarboxylate in an inert solvent, or alternatively by first reacting with a reagent such as, but not limited to, a bromoalkyl chloride or chloroalkyl tosylate to provide an intermediate haloalkoxy group which can be converted to the above described groups by subsequent reaction with an appropriately substituted nucleophile;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is a HOxe2x80x94(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein q and R5 are as defined above, by the reaction in an inert solvent with an alkyl or phenyl substituted isocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90O, or using a base such as pyridine, with a reagent (R5)2NCOCl;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is carboxy or a alkoxycarbonyl group of 2-7 carbon atoms, it can be converted to the corresponding hydroxymethyl group by reduction with an appropriate reducing agent such as lithium borohydride, or lithium aluminum hydride in a inert solvent; the hydroxymethyl group, in turn, can be converted to the corresponding halomethyl group by reaction in an inert solvent with a halogenating reagent such as phosphorus tribromide to give a bromomethyl group, or phosphorus pentachloride to give a chloromethyl group. The hydroxymethyl group can be acylated with an appropriate acid chloride, anhydride, or mixed anhydride in an inert solvent using pyridine or a trialkylamine as a base to give the compounds of this invention with the corresponding alkanoyloxymethyl group of 2-7 carbon atoms, alkenoyloxymethyl group of 2-7 carbon atoms, or alkynoyloxymethyl group of 2-7 carbon atoms;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is a balomethyl group, it can be converted to the corresponding groups: 
wherein R6, R7, R8, R9, M, W, Het, p and q are as hereinabove defined by reacting with the appropriately substituted alcohol, amine or mercaptan in an inert solvent such as dioxane or acetonitrile and a base such as triethylamine or potassium carbonate;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by reacting with the appropriately substituted acid chloride or mixed anhydride (which is prepared from the corresponding carboxylic acid) in an inert solvent such as tetrahydrofuran (THF) in the presence of an organic base such as pyridine, triethylamine or N-methyl morpholine;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by reacting with phosgene in an inert solvent such as toluene in the presence of a base such as pyridine to give an isocyanate which, in turn, is treated with an excess of the alcohol R5xe2x80x94OH;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by the reaction in an inert solvent with an alkyl or phenyl substituted isocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90O, or by reacting with phosgene in an inert solvent such as toluene in the presence of a base such as pyridine to give an isocyanate which, in turn, is treated with an excess of amine (R5)2NH;
where one or more of Q1, Q2, Q3 or Q4 of Formula 44 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by the reaction in an inert solvent with an alkyl or phenyl substituted isothiocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90S, or by reacting with 1,1xe2x80x2-thiocarbonyldiimidazole in an inert solvent such as toluene in the presence of a base such as pyridine to give an isothiocyanate which, in turn, is treated with an excess of amine (R5)2NH.
Intermediate 43 can also be prepared as described below and in Flowsheet 12.
By reacting substituted 2-fluorobenzonitriles of Formula 45 with methyl thioglycolate and a base such as potassium hydroxide or triethylamine in an inert solvent such as dimethyl sulfoxide (DMSO) or aqueous dimethyl formamide (DMF) with or without heating provides compounds of Formula 41. Heating the substituted anilines of Formula 41 with N-methyl piperazine in an inert solvent such as N-methyl pyrrolidine (NMP) provides intermediates 46. Treatment of 46 with ethyl (ethoxymethylene)cyanoacetate gives intermediates 47. Cyclization of 47 in refluxing 1:3 biphenyl/diphenyl ether to provide compounds of Formula 43, or the 4-hydroxy[1]benzothieno[3,2-b]pyridine-3-carbonitriles tautomers thereof, which can be converted to the compounds of this invention using the procedures outlined in Flowsheet 11.
The 2-fluorobenzonitriles of Formula 45 are either commercially available, or are already known to the art or can be prepared by procedures known in the art. 
The preparation of the compounds and intermediates of this invention encompassed by Formula 52 is described below and in Flowsheet 13 where Ar, X, Q1, Q2, Q3 and Q4 and n are as hereinabove defined.
Reaction of substituted 2-nitrophenols of Formula 48 with ethyl bromoacetate and a base such as potassium carbonate in an inert solvent such as dimethyl formamide (DMF) with or without heating, followed by further treatment with potassium t-butoxide in an inert solvent such as tetrahydrofuran (THF) provides compounds of Formula 49. Heating the substituted aniline of Formula 49 with dimethylformamide dimethyl acetal with or without a solvent gives intermediates of Formula 50. The reaction of 50 with from one to ten equivalents of acetonitrile using a base such as n-butyllithium, sodium methoxide or the like in an inert solvent gives the 4-oxo-1,4-dihydro[1]benzofuro[3,2-b]pyridine-3-carbonitrile 51, or the 4-hydroxy[1]benzofuro[3,2-b]pyridine-3-carbonitriles tautomers thereof Heating 51 with or without solvent with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the corresponding 4-chloro[1]benzofuro[3,2-b]pyridine-3-carbonitriles. Condensation of 4-chloro[1]benzofuro[3,2-b]pyridine-3-carbonitriles with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5 gives the benzofuro[3,2-b]pyridine-3-carbonitriles of Formula 52; this condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in alcohol solvents, and the like. In those cases where the Ar and/or Q1, Q2, Q3 and Q4 substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar and/or Q1, Q2, Q3 and Q4 substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 48 where Q1, Q2, Q3 and Q4 contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with ethyl bromoacetate. In those cases, in intermediates 5 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with the 4-chloro[1]benzofuro[3,2-b]pyridine-3-carbonitriles. The same amine or alcohol protecting groups hereinabove described can be used and they can be removed from the products 54 as previously described.
The 2-nitrophenols of Formula 48 are either commercially available, or are already known to the art or can be prepared by procedures known in the art. 
Converting the Q1, Q2, Q3 and Q4 groups to R1, R2, R3 and R4 groups can be accomplished through any conventionally known techniques, for example:
Where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is a nitro group, it can be converted to the corresponding amino group by reduction using a reducing agent such as iron in acetic acid;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is an amino group, it can be converted to the corresponding dialkylamino group of 2 to 12 carbon atoms by alkylation with at least two equivalents of an alkyl halide of 1 to 6 carbon atoms by heating in an inert solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is a methoxy group, it can be converted to the corresponding hydroxy group by reaction with a demethylating agent such as boron tribromide in an inert solvent or by heating with pyridinium chloride with or without solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is an amino group, it can be converted to the corresponding alkylsulfonamido, alkenylsulfonamido, or alkynylsulfonamido group of 2 to 6 carbon atoms by the reaction with an alkylsulfonyl chloride, alkenylsulfonyl chloride, or alkynylsulfonyl chloride, respectively, in an inert solvent using a base such as triethylamine or pyridine;
where two of Q1, Q2, Q3 or Q4 of Formula 52 are contiguous methoxy groups, the corresponding compound with contiguous hydroxy groups can be prepared by using a demethylating agent such as boron tribromide in an inert solvent or by heating with pyridinium chloride with or without solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is an amino group, it can be converted to the corresponding alkylamino group of 1 to 6 carbon atoms by alkylation with one equivalent of an alkyl halide of 1 to 6 carbon atoms by heating in an inert solvent or by reductive alkylation using an aldehyde of 1 to 6 carbon atoms and a reducing agent such as sodium cyanoborohydride in a protic solvent such as water or alcohol, or mixtures thereof;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is hydroxy, it can be converted to the corresponding alkanoyloxy, group of 1-6 carbon atoms by reaction with an appropriate carboxylic acid chloride, anhydride, or mixed anhydride in a inert solvent using pyridine or a trialkylamine as a base;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is hydroxy, it can be converted to the corresponding alkenoyloxy group of 1-6 carbon atoms by reaction with an appropriate carboxylic acid chloride, anhydride, or mixed anhydride in an inert solvent using pyridine or a trialkylamine as a base;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is hydroxy, it can be converted to the corresponding groups: 
wherein V is oxygen, R6, R7, R8, R9, M, W, Het, Ph, p and q are as hereinabove defined and g=2-6 and k=2-4, by reacting with the appropriately substituted alcohol using triphenyl phosphine and diethyl azodicarboxylate in an inert solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is a HOxe2x80x94(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein q and R5 are as hereinabove defined, by the reaction in an inert solvent with an alkyl or phenyl substituted isocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90O, or using a base such as pyridine, with a reagent (R5)2NCOCl;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is carboxy or a alkoxycarbonyl group of 2-7 carbon atoms, it can be converted to the corresponding hydroxymethyl group by reduction with an appropriate reducing agent such as lithium borohydride, or lithium aluminum hydride in an inert solvent; the hydroxymethyl group, in turn, can be converted to the corresponding halomethyl group by reaction in an inert solvent with a halogenating reagent such as phosphorus tribromide to give a bromomethyl group, or phosphorus pentachloride to give a chloromethyl group. The hydroxymethyl group can be acylated with an appropriate acid chloride, anhydride, or mixed anhydride in an inert solvent using pyridine or a trialkylamine as a base to give the compounds of this invention with the corresponding alkanoyloxymethyl group of 2-7 carbon atoms, alkenoyloxymethyl group of 2-7 carbon atoms, or alkynoyloxymethyl group of 2-7 carbon atoms;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is a halomethyl group, it can be converted to the corresponding groups: 
wherein R6, R7, R9, R9, M, W, Het, p and q are as hereinabove defined by reacting with the appropriately substituted alcohol, amine or mercaptan in an inert solvent such as dioxane or acetonitrile and a base such as triethylamine or potassium carbonate;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined by reacting with the appropriately substituted acid chloride or mixed anhydride (which is prepared from the corresponding carboxylic acid) in an inert solvent such as tetrahydrofuran (THF) in the presence of an organic base such as pyridine, triethylamine or N-methylmorpholine;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by reacting with phosgene in an inert solvent such as toluene in the presence of a base such as pyridine to give an isocyanate which, in turn, is treated with an excess of the alcohol R5xe2x80x94OH;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by the reaction in an inert solvent with an alkyl or phenyl substituted isocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90O, or by reacting with phosgene in an inert solvent such as toluene in the presence of a base such as pyridine to give an isocyanate which, in turn, is treated with an excess of amine (R5)2NH;
where one or more of Q1, Q2, Q3 or Q4 of Formula 52 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by the reaction in an inert solvent with an alkyl or phenyl substituted isothiocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90S, or by reacting with 1,1xe2x80x2-thiocarbonyldiimidazole in an inert solvent such as toluene in the presence of a base such as pyridine to give an isothiocyanate which, in turn, is treated with an excess of amine (R5)2NH.
The preparation of the compounds and intermediates of this invention encompassed by Formula 59 is described below and in Flowsheet 14 where Ar, X, Q1, Q2, Q3 and Q4 and n are as hereinabove defined.
Reaction of substituted benzaldehydes (Formula 53) with a nitrating agent such as, but not limited to fuming nitric acid, provides substituted nitrobenzaldehyde intermediates of Formula 54. The condensation reaction of the substituted nitrobenzaldehyde intermediates 54 with methyl cyanoacetate and a base such as piperidine in an alcoholic solvent such as methanol, with or without heating, provides the corresponding substituted 2-cyano-3-(2-nitrophenyl)acrylic acid methyl esters 55. Reduction of the substituted 2-cyano-3-(2-nitrophenyl)acrylic acid methyl esters 55 a reducing agent such as, but not limited to, iron (O) in an alcoholic solvent provides the substituted 2-aminoquinoline-3-carboxylic acid methyl ester intermediates of Formula 56. Heating the substituted 2-aminoquinoline-3-carboxylic acid methyl ester intermediates of Formula 56 with dimethylformamide dimethyl acetal with or without a solvent gives intermediates of Formula 57. The reaction of 57 with from one to ten equivalents of acetonitrile using a base such as n-butyllithium, sodium methoxide or the like in an inert solvent gives the 4-oxo-1,4-dihydrobenzo[b][1,8]naphthyridine-3-carbonitriles 58, or the 4-hydroxy-benzo[b][1,8]naphthyridine-3-carbonitrile tautomers thereof. Heating 58 with or without solvent with a chlorinating agent such as phosphorus oxychloride or oxalyl chloride provides the corresponding 4-chlorobenzo[b][1,8]naphthyridine-3-carbonitriles. Condensation of 4-chlorobenzo[b][1,8]naphthyridine-3-carbonitriles with a nucleophilic amine, aniline, mercaptan, thiophenol, phenol, or alcohol reagent of Formula 5 gives the benzo[b][1,8]naphthyridine-3-carbonitriles of Formula 59; this condensation can be accelerated by heating the reaction mixture together with one equivalent of pyridine hydrochloride or by using bases such as trialkylamines, sodium hydride in an inert solvent, sodium or potassium alkoxides in alcohol solvents, and the like. In those cases where the Ar and/or Q1, Q2, Q3 and Q4 substituents may contain an asymmetric carbon atom, the intermediates can be used as the racemate or as the individual R or S enantiomers in which case the compounds of this invention will be in the racemic or R and S optically active forms, respectively. In cases where the Ar and/or Q1, Q2, Q3 and Q4 substituents may contain more than one asymmetric carbon atoms, diastereomers may be present; these can be separated by methods well known in the art including, but not limited to, fractional crystallization and chromatographic methods. In those cases, in intermediates 54 where Q1, Q2, Q3 and Q4 contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with methyl cyanoacetate. In those cases, in intermediates 5 where Ar contains primary or secondary amino groups or hydroxyl groups, it may be necessary to protect these groups prior to the reaction with the 4-chlorobenzo[b][1,8]naphthyridine-3-carbonitriles. The same amine or alcohol protecting groups hereinabove described can be used and they can be removed from the products 59 as previously described.
The benzaldehydes of Formula 53 are either commercially available, or are already known to the art or can be prepared by procedures known in the art. 
Converting the Q1, Q2, Q3 and Q4 groups to R1, R2, R3 and R4 groups can be accomplished through any conventionally known techniques, for example:
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is a nitro group, it can be converted to the corresponding amino group by reduction using a reducing agent such as iron in acetic acid;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is an amino group, it can be converted to the corresponding dialkylamino group of 2 to 12 carbon atoms by alkylation with at least two equivalents of an alkyl halide of 1 to 6 carbon atoms by heating in an inert solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is a methoxy group, it can be converted to the corresponding hydroxy group by reaction with a demethylating agent such as boron tribromide in an inert solvent or by heating with pyridinium chloride with or without solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is an amino group, it can be converted to the corresponding alkylsulfonamido, alkenylsulfonamido, or alkynylsulfonamido group of 2 to 6 carbon atoms by the reaction with an alkylsulfonyl chloride, alkenylsulfonyl chloride, or alkynylsulfonyl chloride, respectively, in an inert solvent using a base such as triethylamine or pyridine;
where two of Q1, Q2, Q3 or Q4 of Formula 59 are contiguous methoxy groups, the corresponding compound with contiguous hydroxy groups can be prepared by using a demethylating agent such as boron tribromide in an inert solvent or by heating with pyridinium chloride with or without solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is an amino group, it can be converted to the corresponding alkylamino group of 1 to 6 carbon atoms by alkylation with one equivalent of an alkyl halide of 1 to 6 carbon atoms by heating in an inert solvent or by reductive alkylation using an aldehyde of 1 to 6 carbon atoms and a reducing agent such as sodium cyanoborohydride in a protic solvent such as water or alcohol, or mixtures thereof;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is hydroxy, it can be converted to the corresponding alkanoyloxy, group of 1-6 carbon atoms by reaction with an appropriate carboxylic acid chloride, anhydride, or mixed anhydride in a inert solvent using pyridine or a trialkylamine as a base;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is hydroxy, it can be converted to the corresponding alkenoyloxy group of 1-6 carbon atoms by reaction with an appropriate carboxylic acid chloride, anhydride, or mixed anhydride in an inert solvent using pyridine or a trialkylamine as a base;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is hydroxy, it can be converted to the corresponding groups: 
wherein V is oxygen, R6, R7, R8, R9, M, W, Het, Ph, p and q are as hereinabove defined and g=2-6 and k=2-4, by reacting with the appropriately substituted alcohol using triphenyl phosphine and diethyl azodicarboxylate in an inert solvent;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is a HOxe2x80x94(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein q and R5 are as hereinabove defined, by the reaction in an inert solvent with an alkyl or phenyl substituted isocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90O, or using a base such as pyridine, with a reagent (R5)2NCOCl;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is carboxy or a alkoxycarbonyl group of 2-7 carbon atoms, it can be converted to the corresponding hydroxymethyl group by reduction with an appropriate reducing agent such as lithium borohydride, or lithium aluminum hydride in an inert solvent; the hydroxymethyl group, in turn, can be converted to the corresponding halomethyl group by reaction in an inert solvent with a halogenating reagent such as phosphorus tribromide to give a bromomethyl group, or phosphorus pentachloride to give a chloromethyl group. The hydroxymethyl group can be acylated with an appropriate acid chloride, anhydride, or mixed anhydride in an inert solvent using pyridine or a trialkylamine as a base to give the compounds of this invention with the corresponding alkanoyloxymethyl group of 2-7 carbon atoms, alkenoyloxymethyl group of 2-7 carbon atoms, or alkynoyloxymethyl group of 2-7 carbon atoms;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is a halomethyl group, it can be converted to the corresponding groups: 
wherein R6, R7, R8, R9, M, W, Het, p and q are as hereinabove defined by reacting with the appropriately substituted alcohol, amine or mercaptan in an inert solvent such as dioxane or acetonitrile and a base such as triethylamine or potassium carbonate;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined by reacting with the appropriately substituted acid chloride or mixed anhydride (which is prepared from the corresponding carboxylic acid) in an inert solvent such as tetrahydrofuran (THF) in the presence of an organic base such as pyridine, triethylamine or N-methylmorpholine;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by reacting with phosgene in an inert solvent such as toluene in the presence of a base such as pyridine to give an isocyanate which, in turn, is treated with an excess of the alcohol R5xe2x80x94OH;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by the reaction in an inert solvent with an alkyl or phenyl substituted isocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90O, or by reacting with phosgene in an inert solvent such as toluene in the presence of a base such as pyridine to give an isocyanate which, in turn, is treated with an excess of amine (R5)2NH;
where one or more of Q1, Q2, Q3 or Q4 of Formula 59 is a H2N(CH2)qxe2x80x94 group, it can be converted to the corresponding groups: 
wherein R5 and q are as hereinabove defined, by the reaction in an inert solvent with an alkyl or phenyl substituted isothiocyanate, R5xe2x80x94Nxe2x95x90Cxe2x95x90S, or by reacting with 1,1xe2x80x2-thiocarbonyldiimidazole in an inert solvent such as toluene in the presence of a base such as pyridine to give an isothiocyanate which, in turn, is treated with an excess of amine (R5)2NH.
Compounds of this invention are evaluated in several standard pharmacological test procedures that showed that the compounds of this invention possess significant activity as inhibitors of protein kinases and are antiproliferative agents. Among the disease states which can be treated or inhibited by protein kinase inhibitors include those in which the etiology is at least in part caused by a defect upstream in a signaling pathway from a protein kinase (e.g., colon cancer); those in which the etiology is at least in part caused by an overexpressed protein kinase (e.g., lung cancer and colonic polyps); and those in which the etiology is at least in part caused by a dysregulated protein kinase (gene turned on at all times; glioblastoma).
Based on the activity shown in the standard pharmacological test procedures, the compounds of this invention are therefore useful as antineoplastic agents. In particular, these compounds are useful in treating, inhibiting the growth of, or eradicating neoplasms such as those of the breast, kidney, bladder, mouth, larynx, esophagus, stomach, colon, ovary, lung, pancreas, liver, prostate and skin.
In addition to having antineoplastic properties, the compounds of the present invention are useful in treating or inhibiting a variety of protein tyrosine kinase-associated disorders including: polycystic kidney disease, colonic polyps, restenosis; atherosclerosis; angiofibromas; hemangiomas; diabetes; acute and chronic nephropathies; Kaposi""s sarcoma; neovascularization associated with macular degeneration; rheumatoid arthritis; osteoarthritis; transplant rejection; psoriasis; lupus; graft versus host disease; glomerulonephritis; respiratory and skin allergies; autoimmune alopecia; Autoimmune Hyperthyroidism; multiple sclerosis; atopic dermatitis; and systemic sclerosis; and are useful as antibacterial and antiviral agents.
As used in accordance with this invention, the term providing an effective amount of a compound means either directly administering such compound, or administering a prodrug, derivative, or analog which will form an effective amount of the compound within the body.
The test procedures used and results obtained are shown below.
Inhibition of Epidermal Growth Factor Receptor Kinase (EGF-R ) Using Recombinant Enzyme
Representative test compounds are evaluated in a standard pharmacological test procedure to measure their ability to inhibit the phosphorylation of the tyrosine residue of a peptide substrate catalyzed by the enzyme epidermal growth factor receptor kinase. The peptide substrate (RRxe2x80x94SRC) has the sequence arg-arg-leu-ile-glu-asp-ala-glu-tyr-ala-ala-arg-gly. The enzyme used in this assay is the His-tagged cytoplasmic domain of EGFR. A recombinant baculovirus (vHcEGFR52) is constructed containing the EGFR cDNA encoding amino acids 645-1186 preceded by Met-Ala-(His)6. Sf9 cells in 100 mm plates are infected at a multiplicity of infection of 10 pfu/cell and cells are harvested 48 h post infection. A cytoplasmic extract is prepared using 1% Triton X-100 and applied to Nixe2x80x94NTA column. After washing the column with 20 mM imidazole, HcEGFR is eluted with 250 mM imidazole (in 50 mM Na2HPO4, pH 8.0, 300 mM NaCl). Fractions collected are dialyzed against 10 mM HEPES, pH 7.0, 50 mM NaCl, 10% glycerol, 1 ug/mL antipain and leupeptin and 0.1 mM Pefabloc SC. The protein is frozen in dry ice/methanol and stored xe2x88x9270xc2x0 C.
Test compounds are made into 10 mg/mL stock solutions in 100% dimethylsulfoxide (DMSO). Prior to experiment, stock solutions are diluted to 500 uM with 100% DMSO and then serially diluted to the desired concentration with HEPES buffer (30 mM HEPES pH 7.4).
For the enzyme reaction, 10 uL of each inhibitor (at various concentrations) are added to each well of a 96-well plate. To this is added 3 uL of enzyme (1:10 dilution in 10 mM HEPES, pH 7.4 for final conc. of 1:120). This is allowed to sit for 10 min on ice and is followed by the addition of 5 ul peptide (80 uM final conc.), 10 ul of 4xc3x97 Buffer containing 50 mM HEPES (pH 7.4), 200 mM Na3VO4, 40 mM MnCl2, 80 uM ATP, 0.25 uL 33P-ATP ( greater than 2500 Ci/mmol; Amersham) and 12 uL H2O. The reaction is allowed to run for 90 min at room temperature and is followed by spotting the entire volume onto precut P81 filter papers. The filter discs are washed 2xc3x97 with 0.5% phosphoric acid and radioactivity is measured using a liquid scintillation counter.
The inhibition data for representative compounds of the invention are shown below in TABLE 1. The IC50 is the concentration of test compound needed to reduce the total amount of phosphorylated substrate by 50%. The % inhibition of the test compound is determined for at least three different concentrations and the IC50 value is evaluated from the dose response curve. The % inhibition is evaluated with the following formula:
% inhibition=100xe2x88x92[CPM(drug)/CPM(control)]xc3x97100
where CPM(drug) is in units of counts per minute and is a number expressing the amount of radiolabled ATP (xcex3-33P) incorporated onto the RR-SRC peptide substrate by the enzyme after 90 minutes at room temperature in the presence of test compound as measured by liquid scintillation counting. CPM(control) is in units of counts per minute and is a number expressing the amount of radiolabled ATP (xcex3-33P) incorporated into the RR-SRC peptide substrate by the enzyme after 90 minutes at room temperature in the absence of test compound as measured by liquid scintillation counting. The CPM values are corrected for the background counts produced by ATP in the absence of the enzymatic reaction.
Inhibition of Kinase Insert Domain Containing Receptor (KDR; the Catalytic Domain of the VEGF Receptor)
KDR protein is mixed, in the presence or absence of a inhibitor compound, with a substrate peptide to be phosphorylated (a copolymer of glutamic acid and tyrosine, E:Y::4:1) and other cofactors such as Mg++ and sodium vanadate (a protein tyrosine phosphatase inhibitor) in an appropriate buffer to maintain pH (7.2). ATP and a radioactive tracer (either P32- or P33-labeled ATP) is then added to initiate phosphorylation. After incubation, the radioactive phosphate associated with the acid-insoluble fraction of the test procedure mixture is then quantified as reflection of substrate phosphorylation. This radioactive format is used to identify inhibitors of KDR tyrosine kinase activity where the IC50 is the concentration of drug that inhibits substrate phosphorylation by 50%. The results obtained for representative compounds of this invention are listed in Table 2.
Mitogen Activated Protein Kinase (MAPK) Test Procedure
To evaluate inhibitors of the MAP (mitogen activated protein) kinase a two component coupled standard pharmacological test procedure, which measures phosphorylation of a serine/threonine residue in an appropriate sequence in the substrate in the presence and absence of a putative inhibitor, is used. Recombinant human MEK 1 (MAPKK) is first used to activate recombinant human ERK2 (MAPK) and the activated MAPK (ERK) is incubated with substrate (myelin basic protein peptide (MBPP) or Myc peptide) in the presence of ATP, Mg+2 and radiolabeled 33P ATP. The phosphorylated peptide is captured on a P 81 phosphocellulose filter (paper filter or embedded in microtiter plate) washed and counted by scintillation methods.
The peptide substrates used in the assay are MBPP, peptide substrate (APRTPGGRR), or synthetic Myc substrate, (KKFELLPTPPLSPSRRxe2x80xa25 TFA). The recombinant enzymes used are prepared as GST fusion proteins of human ERK 2 and human MEK 1. Inhibitor samples are prepared as 10xc3x97stocks in 10% DMSO and an appropriate aliquot is used to deliver either 10 ug/ml for a single point screening dose or 100 to 0.0001 uM final concentration for a dose response curve. Final DMSO concentrations are less than or equal to 1%.
The reaction is run as follows in 50 mM Tris kinase buffer, pH 7.4 in a reaction volume of 50 ul. The appropriate volume of kinase buffer and inhibitor sample is added to the tube. Appropriate dilution of enzyme is delivered to give 2-5 ug recombinant MAPK (Erk) per tube. The inhibitor is incubated with MAPK (Erk) for 30 min at 0 deg. C. Recombinant Mek (MAPKK) (0.5-2.5 ug) or fully activated Mek (0.05-0.1 units) is added to activate the Erk and incubated for 30 min at 30xc2x0 C. Then substrate and xcex333P ATP are added to give a final concentration of 0.5-1 mM MBPP or 250-500 uM Myc; 0.2-0.5 uCi gamma P 33 ATP/tube; 50 xcexcM ATP final concentration. Samples are incubated at 30xc2x0 C. for 30 minutes and the reaction is stopped by adding 25 xcexcl of ice cold 10% trichloroacetic acid (TCA). After samples are chilled on ice for 30 min, 20 xcexcl of sample is transferred onto P 81 phosphocellulose filter. Filter papers are washed 2 times with a large volume of 1% acetic acid, then 2 times with water. The filters are briefly air dried before addition of scintillant and samples are counted in the appropriate scintillation counter set up for reading 33P isotope. Samples include a positive control (activated enzyme plus substrate); a no enzyme control; a no substrate control; samples with different concentrations of putative inhibitor; and samples with reference inhibitors (other active compounds or non-specific inhibitors such as staurosporine or K252 B).
The raw data is captured as cpm. Sample replicates are averaged and corrected for background count. Mean cpm data is tabulated by group and % inhibition by a test compound is calculated as (corrected cpm control-corrected. cpm sample/control)xc3x97100=% inhibition). If several concentrations of inhibitor are tested, IC50 values (the concentration which gives 50% inhibition) are determined graphically. The results obtained for representative compounds of this invention are listed in Table 2.
Src Kinase Test Procedrue
Inhibitors of p60c-src (partially purified preparation purchased from Upstate Biotechnologies) tyrosine kinase activity are analyzed in an Elisa format. The Boehringer Mannheim Tyrosine Kinase Assay Kit (Catalog number 1-534505) with a cdc2 substrate peptide containing Tyr15 is used for the assay. HRP-conjugated anti-phosphotyrosine is used to detect phosphorylated peptide via a color reaction. Conditions recommended by the manufacturer are employed.
Reaction conditions: Five microliter aliquots of each compound prepared fresh at the time of the assay are added as a solution in 10 mM HEPES pH 7.5, 10% DMSO to the reaction well. Thirty-five microliters of reaction mix containing Src, buffer and peptide/bovine serum albumin mix are added to the compound wells and incubated at 30xc2x0 C. for 10 minutes (reaction buffer: 50 mM TrisHCl pH 7.5, 10 mM MgCl2, 0.1 mM EGTA, 0.5 mM Na3VO4). The reaction is started by addition of 10 microliters of ATP, incubated at 30xc2x0 C. for 1 hour, and stopped by addition of 20 microliters of 0.5M EDTA. The reaction mixture with the phosphorylated peptide is then transferred to a streptavidin-coated microtiter plate (provided in the kit) and allowed to bind for 20 minutes. Unbound peptide and reaction mixture is decanted and the plate is washed with PBS six times. Horseradish peroxidase-conjugated phosphotyrosine antibody supplied in the kit is incubated with the plate for one hour, then decanted. The plate is again washed with PBS six times. Substrate (provided in the kit) is added and absorbance at 405 nm is measured.
Activity is determined as % inhibition as calculated by the formula:
(1xe2x88x92Abs/Abs(max))xc3x97100=% inhibition.
Where multiple concentrations of the test agent are used, an IC50 (concentration which gives 50% inhibition) could be determined.
The results obtained for representative compounds of this invention are listed in Table 2.
Cell Proliferation Test Procedure
HT-29 cells: Compound effectiveness at inhibiting cell proliferation on plastic is performed in a 96-well format by plating 5000 cells per well in appropriate medium on day one, followed by compound addition on day 2 in serial two-fold dilutions. On day five, compound is washed away and medium containing MTS reagent (Promega) is added. Relative cell number is determined by reading the absorbance at 490 nm of a dye produced by an NAD-dependent cellular enzymatic reaction. These data are shown below in Table 3.
Anchorage Independent Src-transformed Fibroblast Proliferation Test Procedure: Rat2 fibroblasts stably transformed with a plasmid containing a CMV promotor controlled v-Src/Hu c-Src fusion gene in which the catalytic domain of human c-Src is inserted in place of the v-Src catalytic domain in the v-Src gene are used for the measurement of src dependent suspension growth. Ultra-low cluster plates (Costar #3474) are seeded with 10,000 cells per well on Day 1. Compound is added in serial two-fold dilutions from 10 micromolar to 0.009 micromolar on Day 2 and MTS reagent (Promega) is added on Day 5 (100 microliters of MTS/medium mix+100 microliters of medium already on the cells and the absorbance is measured at 490 nm. The results are analyzed as follows to yield an IC50 for proliferation (micromolar units) as follows: % inhibition=(Abs490 nm sample-blank)/(Abs490 nm no cmpd control-blank)xc3x97100%. These data are shown below in Table 3.
Inhibition of Cancer Cell Growth as Measured by Cell Number
Human tumor cell lines are plated in 96-well plates (250 xcexcl/well, 1-6xc3x97104 cells/ml) in RPMI 1640 medium, containing 5% FBS (Fetal Bovine Serum). Twenty four hours after plating, test compounds are added at various concentrations. After 48 hours exposure to test compounds, cells are fixed with trichloroacetic acid, and stained with Sulforhodamine B. After washing with trichloroacetic acid, bound dye is solubilized in 10 mM Tris base and optical density is determined using plate reader. Under conditions of the assay the optical density is proportional to the number of cells in the well. IC50s (concentrations causing 50% inhibition of cell growth) are determined from the growth inhibition plots. The test procedure is described in detail by Philip Skehan et. al, J. Natl. Canc. Inst., 82, 1107-1112 (1990). These data are shown below in Table 4. Information about some of the cell lines used in these test procedures is available from the American Type Tissue Collection: Cell Lines and Hybridomas, 1994 Reference Guide, 8th Edition.
Raf1 Kinase Cascade Assay Procedure
Raf-1 (c-Raf) is used to phosphorylate and activate inactive GST-MEK1 which then can phosphorylate and activate inactive p42 GST-MAPK, which subsequently is measured for phosphorylation of the TEY sequence (aa""s 202-204) by a phospho-specific antibody from Sigma (cat. #77439219041) Reagents: Sf9 insect cell lysate containing full length 6his-tagged recombinant human c-Raf. (Specific Activity: xcx9c200 U/ml). Human Non-active Mek-1-GST and human GST-MAP kinase (recombinant proteins produced in E. coli).
Stock Solutions Raf Assay
1. Assay Dilution Buffer (ADB): 20 mM MOPS, pH 7.2, 25 mM xcex2-glycerol phosphate, 5 mM EGTA, 1 mM sodium orthovanadate, 1 mM dithiothreitol.
2. Magnesium/ATP Cocktail: 500 xcexcM cold ATP and 75 mM magnesium chloride in ADB.
4. Active Kinase: Human Active c-Raf: Use at 0.4 U per assay point.
5. Non-active GST-MEK1: Use at 0.1 xcexcg per assay point.
6. Non-active GST-p42 MAP Kinase: Use at 1.0 xcexcg per assay point.
Stock Solutions ELISA
1. TBSTxe2x80x94Tris (50 mM, pH 7.5), NaCl (150 mM), Tween-20 (0.05%)
2. Superblock (Pierce)
3. Anti-GST Ab (Pharmacia)
4. Anti-Phospho MAPK (Sigma)
5. Anti-Mouse Ab/Europium conjugate (Wallac)
Assay Procedure
First Stage: c-Raf Dependent Activation of GST-MEK and GST-MAPK
1. Add 20 ml of ADB per assay (i.e. per well of a 96 well plate)
2. Add 10 ml of 0.5 mM cold ATP and 75 mM magnesium chloride in ADB.
3. Add 2 ml of c-Raf (0.4 U/assay), in conjunction with 1.6 ml non-active MEKI (0.4 mg/assay).
4. Add 4 ml of non-active GST-p42 MAP Kinase (1.0 mg/assay).
5. Incubate for 60 minutes at 30xc2x0 C. in a shaking incubator.
6. Transfer this mixture to an anti-GST Ab coated 96 well plate (Nunc Immunosorb plates coated o/n with a-GST, then blocked with Pierce Superblock).
7. Incubate for 60 minutes at 30xc2x0 C. in a shaking incubator Wash 3xc3x97 with TBST, add Anti-Phospho MAPK (Sigma) (1:3000)
6. Incubate for 60 minutes at 30xc2x0 C. in a shaking incubator
7. Wash 3xc3x97 with TBST, add Anti-Mouse Ab/Europium conjugate (Wallac) (1:500)
8. Incubate for 60 minutes at 30xc2x0 C. in a shaking incubator
9. Wash 3xc3x97 with TBST, Read plates in Wallac Victor model Plate Reader.
10. Collect data analyze in Excel for single point and IC50 determinations.
Single point assayxe2x80x94% inhibition at 10 mg/ml (% Inhibition=1xe2x88x92cpd.treated sample/untreated control). IC50 determinationsxe2x80x94done on compounds from single point assays with  greater than 80% inhibition. Typically Raf-1 assay is run at compound concentrations from 10 xcexcM to 1 nM in half log dilutions. (% inhibition is determined for each compound concentration). The results obtained for representative compounds of this invention are listed in Table 2.
Cell Based Screen for Inhibitors of Raf Kinase
Materials
Cell Lines: Human adenocarcinoma cell line LoVo which is known to be growth inhibited by low nM concentrations of a reference standard inhibitor of Ras and human adenocarcinoma cell line CaCo-2, which is known to be growth resistant to the same reference compound.
Cell Media: RPMI 1640 with 10% Fetal Bovine Serum supplemented with L-glutamine and Pennicilin/Streptomycin.
Compounds: Supplied usually as a 10 mM stock in 100% DMSO.
Normal Saline: 150 mM NaCl
Trichloroacetic Acid (TCA): 50% (w/v) in water
Sulforhodamine B (SRB): 0.4% (w/v) in 1% Acetic Acid
Tris Base: 10 mM in water
Methods
Cells are plated at 2000 cells per well for cell line LoVo and 1500 cells for cell line CaCo-2 in 96 well plates. Cells are plated in media (200 xcexcl) and allowed to adhere overnight at 37xc2x0 C. At 24 hours post plating, compounds are added directly at a volume of 0.5 xcexcl. For the qualitative screen (compounds screened at 25 xcexcM) compound is added directly to cells. For the quantitative screen, compound is first diluted in DMSO to generate concentrations of compound or reference standard of: 1,5, 10 and 25 xe2x96xa1M. It is advisable to make the dilutions in an identical 96 well plate so that compounds can be added using a multichannel micropipettor set at 0.5 xcexcl. The cells are then incubated for four days after which the media is removed using a 12 well manifold by first tipping the plate forward at a 45 degree angle and then inserting the manifold in an upright orientation to prevent the tips of the manifold from disturbing cells at the bottom of the plate. 200 xcexcl of normal saline is then added to each well using an 8 well multichannel pipettor, followed by the careful addition of 50 xcexcl of 50% TCA. The plates are then incubated for 2 hours at 4xc2x0 C. , after which the supernatant is removed using the same technique as above and the plated washed twice with 200 xcexcl water. The plates are then air dried and 50 xcexcl of SRB stock solution is carefully added so that the entire bottom of each well is covered. This again can be used using an 8 well multichannel pipettor. The SRB is incubated with fixed cells for 15 minutes at room temperature after which the SRB is removed with the manifold as described above and the plates washed twice with 350 xcexcl of 1% acetic acid per well each time. The plates are then air dried after which the bound SRB is released from protein by the addition of 200 xcexcl of Tris base. Resolubilizing the SRB is aided by placing the plates on a rotator for 15-30 minutes. The absorbance of each well is determined at 550 or 562 nm using a microtiter plate reader.
Each compound or dilution thereof is performed in triplicate. Outliers are identified by visual inspection of the data. Each plate should have a xe2x80x9c0xe2x80x9d control (vehicle only). Qualitative screen: To calculate % inhibition of a compound at 25 xcexcM, the following formula is used: 1xe2x88x92(experimental absorbance @ 25 xcexcM compound/xe2x80x9c0xe2x80x9d control absorbance)xc3x97100=% inhibition at 25 xcexcM. Compounds having  greater than 50% inhibition at 25 xcexcM are placed in the quantitative assay.
Quantitative Assay: A standard curve is constructed by plotting the concentration of compound against the average absorbance calculated at that concentration. A curve is plotted and the concentration at which the curve passes through the 50% the absorbance mark seen in the xe2x80x9c0xe2x80x9d control well is the IC50 calculated for that compound. Multiple entries for a given compound indicate that it is tested multiple times. The results obtained for representative compounds of this invention are listed in Table 5.
The results shown in tables 1, 2, 3, 4 and 5 demonstrate that the compounds of this invention are potent inhibitors of protein kinases, and are useful as described above.
The compounds of this invention may be formulated neat or may be combined with one or more pharmaceutically acceptable carriers for administration. For example, solvents, diluents and the like, and may be administered orally in such forms as tablets, capsules, dispersible powders, granules, or suspensions containing, for example, from about 0.05 to 5% of suspending agent, syrups containing, for example, from about 10 to 50% of sugar, and elixirs containing, for example, from about 20 to 50% ethanol, and the like, or parenterally in the form of sterile injectable solution or suspension containing from about 0.05 to 5% suspending agent in an isotonic medium. Such pharmaceutical preparations may contain, for example, from about 0.05 up to about 90% of the active ingredient in combination with the carrier, more usually between about 5% and 60% by weight.
The effective dosage of active ingredient employed may vary depending on the particular compound employed, the mode of administration and the severity of the condition being treated. However, in general, satisfactory results are obtained when the compounds of the invention are administered at a daily dosage of from about 0.5 to about 1000 mg/kg of animal body weight, optionally given in divided doses two to four times a day, or in sustained release form. For most large mammals the total daily dosage is from about 1 to 1000 mg, preferably from about 2 to 500 mg. Dosage forms suitable for internal use comprise from about 0.5 to 1000 mg of the active compound in intimate admixture with a solid or liquid pharmaceutically acceptable carrier. This dosage regimen may be adjusted to provide the optimal therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation.
The compounds of this invention may be administered orally as well as by intravenous, intramuscular, or subcutaneous routes. Solid carriers include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose and kaolin, while liquid carriers include sterile water, polyethylene glycols, non-ionic surfactants and edible oils such as corn, peanut and sesame oils, as are appropriate to the nature of the active ingredient and the particular form of administration desired. Adjuvants customarily employed in the preparation of pharmaceutical compositions may be advantageously included, such as flavoring agents, coloring agents, preserving agents, and antioxidants, for example, vitamin E, ascorbic acid, BHT and BHA.
The preferred pharmaceutical compositions from the standpoint of ease of preparation and administration are solid compositions, particularly tablets and hard-filled or liquid-filled capsules. Oral administration of the compounds is preferred.
In some cases it may be desirable to administer the compounds directly to the airways in the form of an aerosol.
The compounds of this invention may also be administered parenterally or intraperitoneally. Solutions or suspensions of these active compounds as a free base or pharmacologically acceptable salt can be prepared in water suitably mixed with a surfactant such as hydroxy-propylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof in oils. Under ordinary conditions of storage and use, these preparation contain a preservative to prevent the growth of microorganisms.
The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.
For the treatment of cancer, the compounds of this invention can be administered in combination with other antitumor substances or with radiation therapy. These other substances or radiation treatments can be given at the same or at different times as the compounds of this invention. These combined therapies may effect synergy and result in improved efficacy. For example, the compounds of this invention can be used in combination with mitotic inhibitors such as taxol or vinblastine, alkylating agents such as cisplatin or cyclophosamide, antimetabolites such as 5-fluorouracil or hydroxyurea, DNA intercalators such as adriamycin or bleomycin, topoisomerase inhibitors such as etoposide or camptothecin, antiangiogenic agents such as angiostatin, and antiestrogens such as tamoxifen.
The preparation of representative examples of the compounds of this invention is described below.